Current Biotechnology

Opportunities and Challenges for the Industrial Application of Transgenic Insect-resistant and Herbicide-tolerant Maize in China
Min LI, Lei WANG, Junjie ZOU
2023, 13(2): 157-165. DOI: 10.19586/j.2095-2341.2022.0183
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Maize is an important food and forage crop in China， and the prevention and control of insect pests and weeds is a major bottleneck problem faced by maize production in China. The application of transgenic insect-resistant and herbicide-tolerant maize can reduce the usage of pesticides and play an important role in improving corn yield and harvest quality. Since the commercial application of transgenic insect-resistant and herbicide-tolerant maize abroad in 1996， the lepidoptera pests such as corn borer and fall armyworm have been effectively controlled， and the weeding cost has been reduced， with remarkable economic， social and ecological benefits. Here， the development status of transgenic insect-resistant and herbicide-tolerant maize in the world and China in the past ten years was summarized， the opportunities and challenges faced by the industrialization of transgenic insect-resistant and herbicide-tolerant maize in China were analyzed， and some suggestions were provided for accelerating the industrialization application of transgenic maize in China.

Application Progress of Cellulase in Tobacco
Jie HAO, Xuanwen LI, Bao ZHANG, Chao ZHENG, Zhikang SUN, Qiang JI, Na WU, Han WU, Liqun LI
2023, 13(2): 166-173. DOI: 10.19586/j.2095-2341.2022.0127
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The quality of cigarette raw materials is the basis to determine the cigarettes quality. The main components of cigarette raw materials such as cellulose， pectin， sugar and nicotine are different in different regions， which restricts the development of industrial enterprises. In order to solve the uneven raw materials in different regions， progress has been made in the degradation of macromolecules （such as cellulose） and chemical components （such as nicotine） by biological enzyme technology. In this paper， the research progress of cellulase and its compound enzyme preparation in tobacco was summarized， and the influence of cellulase on tobacco leaf， stem， reconstituted tobacco leaf and tobacco extract was analyzed， in order to provide reference for the production of tobacco leaf and cigarette by biological enzyme.

Research Progress of Cyanobacteria Cell Factories
Junkui ZHAO, Yongzhong LU
2023, 13(2): 174-180. DOI: 10.19586/j.2095-2341.2022.0203
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With strong carbon fixation capacity and amenability to genetic manipulations， cyanobacteria are considered to be ideal cell factories for developing green biotechnology products. However， many research achievements are still in the laboratory stage， and the production efficiency of various chemicals is low for a long time. Combining the advantages of cyanobacteria and making full use of modern biotechnology will help promote the in-depth research and development of cyanobacterial cell factories. Taking the development process of cyanobacterial biotechnology as the main line， this paper introduced the achievements of genetic engineering， metabolic engineering and synthetic biology in the research of cyanobacterial cell factory， and pointed out that "drug and food homology" was the direction of foreign protein expression by cyanobacteria for application. To develop advanced synthetic biological tools so as to transform， design and reconstruct the metabolic pathways of cyanobacteria chassis cells was the only way to achieve efficient production of chemicals. This paper was expected to provide a reference for the in-depth study of cyanobacterial biotechnology.

Progress of Fungal Secondary Metabolites with Potential Herbicidal Activity
Kainan SONG, Linan XIE, Yuquan XU
2023, 13(2): 181-194. DOI: 10.19586/j.2095-2341.2022.0141
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In agricultural production， the widespread use of chemical herbicides， while securing the crop production， has caused many environmental problems and the evolution of herbicide resistance. Therefore， herbicides with more safety and novel modes of action are an urgent need. Fungi can produce a variety of bioactive secondary metabolites， some of which have strong herbicidal activity. In this review， some secondary metabolites isolated from fungi and their herbicidal activities in the past ten years were reviewed according to their structures. The prospect of development and application of fungal herbicidal compounds was discussed，which was expected to provide reference for the research and development of new herbicide.

Research Progress of Nucleic Acid Reference Materials of Food-borne Pathogenic Bacteria
Enze CHENG-CHEN, Minghong JIA, Yueying LI
2023, 13(2): 195-200. DOI: 10.19586/j.2095-2341.2022.0142
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With the rapid development of nucleic acid detection kit in the field of rapid microbial detection， the detection process has the advantages of simple operation， low cost and convenience for on-site detection. In the preparation process of nucleic acid detection kit for food-borne pathogenic bacteria， it is necessary to ensure its qualitative and quantitative measurement ability through metrological traceability. The application of nucleic acid reference materials （NARMs） runs through many fields， such as clinical hygiene， food， instrument verification/calibration， nucleic acid detection， etc. By segmenting genetic materials， the biological safety can be guaranteed to the greatest extent， and the requirements of quality evaluation and metrological traceability can be met. It can effectively control the quality of nucleic acid detection kits and provide an accurate “yardstick” for the quantitative evaluation of nucleic acid detection kits. This paper introduced the development method of NARMs in China， focusing on the current research status of food-borne pathogens and its application in the national standard material resource sharing platform， and analyzed the technical challenges and the possibility of the development of nucleic acid metrology in the future， so as to provide reference ideas for the early diagnosis and prevention of nucleic acid detection industry.

Application of Virus Like Particles Assembled in Vitro in Vaccine and Drug Delivery
Yanyi LI, Na LYU, Sining JIA, Yonghong ZHANG, Hongxia ZHANG, Weiting ZHANG, Jinli CHEN
2023, 13(2): 201-209. DOI: 10.19586/j.2095-2341.2022.0108
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Virus like particles （VLPs） have been widely used in the field of vaccines and therapeutic drug delivery systems in recent years. Compared with VLPs assembled in vivo， VLPs assembled in vitro have many advantages， such as clear composition of assembled oligomers， controllable reaction conditions， avoiding the introduction of host components， and realizing the encapsulation and delivery of drug active components. This review mainly described the production methods of extracellular VLPs， the influencing factors of VLPs formation， and the characterization methods of VLPs assembled in vitro. The application of VLPs assembled in vitro in vaccine and therapeutic drug delivery was also reviewed. It is expected to provide reference ideas for the development and progress of VLPs assembled in vitro assembly technology， and promote VLPs assembled in vitro assembly technology to play a greater role in disease prevention and treatment.

Research Progress on Production and Application of Mannosylerythritol Lipids
Siyuan QIU, Jingxue XU, Yuyang DUAN, Jinyu ZHAO, Wenjing ZHAO, Lixin ZHANG, Guoling REN
2023, 13(2): 210-219. DOI: 10.19586/j.2095-2341.2022.0131
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As　a type of biosurfactant， mannosylerythritol lipids （MELs） have merits with biodegradability， low toxicity and good biocompatibility. Due to its distinct structural， metabolic， and synthetic features， it also performs a wide range of other tasks， including skin restoration， gene transfection， and broad-spectrum antibacterial activity. MELs has a promising future in a number of industries， including medicine， daily chemicals， food， agriculture， and ecological restoration， which is widely recognized as one of the most promising biosurfactants. However， because of the great structural heterogeneity， backward manufacturing style of MELs generated by various species， hazy synthesis and uncertain mechanism of action， it is hard to carry out large-scale commercial production. The paper aimed to clarify the diversity of its structure and function， so as to provide reference for the realization of customized production targeting MELs. By focusing on the aspects of structural characteristics， production purification， application path and so on， the paper was hoped to help reduce the production cost and accelerate the realization of its large-scale application.

Progress on Lipase Immobilization and its Application in Pharmaceutical Synthesis
Yamei ZHOU, Jia LIU, Dan LU, Qingxin KONG
2023, 13(2): 220-227. DOI: 10.19586/j.2095-2341.2022.0207
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As a commonly used biocatalyst， lipase is widely used in pharmaceutical， food and biochemical applications. However， the application of lipase in enzyme catalysis industry was limited because of its poor stability of free lipase， susceptibility to the environment and poor reusability. In response to the shortcomings of free lipase in the field of catalysis， enzyme immobilization technology was developed. The immobilization of lipases has greatly improved their original catalytic activity and stability and has led to a wider range of industrial applications. The use of immobilized lipases with their own excellent properties selectively catalyses the synthesis of the desired product with mild reaction conditions， high yields and few side reactions. The research and applications of lipase immobilization and its use in pharmaceutical synthesis were reviewed and the future of immobilized lipases was given， which was expected to provide reference for the application of immobilized lipase in industry.

Overview of the Main Molecular Mechanisms of Biological Aging
Wei ZHANG, Hongfang WANG, Baohua XU
2023, 13(2): 228-233. DOI: 10.19586/j.2095-2341.2022.0170
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Aging is a normal natural law including physical aging and pathological aging. Like other biological processes， it is regulated by some signaling pathways and molecular mechanisms. Through the research on the mechanism of regulating biological aging， it is found that the signaling pathways interact with each other. This paper reviewed three classical signal pathways related to autophagy and delaying aging， including insulin pathway， rapamycin pathway and Sirtuins family， and summarized the main causes and ways of oxidative stress， cell aging and immune aging affecting aging， hoping to discover new interaction pathways and explore more novel molecular mechanisms and methods to prevent， delay or reduce a variety of diseases related to aging.

Research Progress on the Protective Effects of Hydrogen Gas on Traumatic Brain Injury
Jingyi ZHANG, Xue JIANG, Siyu MA, Zhichao FENG, Yang YI, Chen MA, Yifei SONG, Fei XIE
2023, 13(2): 234-239. DOI: 10.19586/j.2095-2341.2022.0202
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Traumatic brain injury （TBI） is a very common disease in daily trauma， and there is still a lack of drugs with definite clinical interventions， so it is important to develop new treatments for TBI. Hydrogen has been proved to have antioxidant， anti-inflammatory， anti-apoptosis and other effects， and has shown a good protective effect in a variety of diseases. In the past decade， many studies have preliminarily confirmed the protective effect of hydrogen on TBI， and the mechanism underlying the effect of hydrogen has also been investigated. The review focused on the protective effect of different hydrogen intervention methods on TBI and the mechanism underlying the effect of hydrogen， and also put forward the deficiencies in the current research and the direction of future research. The paper was hoped to provide reference and help for clinical and basic researchers of hydrogen.

Research Progress of Nrf2 in Ferroptosis After Spinal Cord Injury
Jingyu CAO, Chengmei LIU, Chenxu QI, Kaiyan DU, Meng CHEN, Siwei HOU
2023, 13(2): 240-246. DOI: 10.19586/j.2095-2341.2022.0178
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Ferroptosis is one of the important pathological mechanisms of neuronal cell injury after spinal cord injury （SCI）. It has received widespread attention at home and abroad in recent years， but no effective breakthrough has been achieved. In recent years， it has been found that oxidative stress plays an important role in the pathophysiological process. Nuclear factor E2-related factor 2 （Nrf2） has become a hot topic of research because of its anti-oxidative stress effect and involvement in neural repair after SCI， but its mechanism of regulating the development of ferroptosis is still unclear. In this paper， we briefly described the basic structure of Nrf2 and its downstream target genes， introduced the mechanism of ferroptosis and explored the potential role of Nrf2 in SCI treatment， and provided ideas for further study of its relationship with iron death in the development of SCI and explore new strategies for SCI treatment.

Cascade Catalysis of Glycosyltransferase and Sucrose Synthase to Produce Astragalin
Zhekang JIA, Xinran CHENG, Wenfang DOU
2023, 13(2): 247-256. DOI: 10.19586/j.2095-2341.2022.0212
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Astragalin is a flavonoid 3-O-glycoside compound with various pharmacological activities such as antioxidant， antiviral and antibacterial. In this study， glucosyltransferase from grapefruit and sucrose synthase from soybean were used to convert kaferol to astragalin， while the glycosylation byproduct uridine diphosphate （UDP） was regenerated to UDP-glucose. Through fed-batch operation， the yield of astragalin reached 1 550.1 mg·L^-1， the conversion rate was 86.5%， and the UDP cycle was 34.8 times. After isolation and purification， the results were basically consistent with the literature， and the reaction product was indeed astragalin. The synthesis method of astragalin proposed in this study greatly reduced the reaction cost and provided an new idea for the industrial production of astragalin.

Research on the Function of CXCL2 During Megakaryocyte Differentiation
Di WANG, Yanhong ZHAO, Yipeng LIANG, Haoze SONG, Lihong SHI, Jingyuan TONG
2023, 13(2): 257-263. DOI: 10.19586/j.2095-2341.2022.0217
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Clinically， the conflict between supply and demand of platelet transfusion is becoming increasingly acute， and research related to platelet generation in vitro has received extensive attention globally， mainly focusing on stem cell sources， cytokines， transcription factors， and turbulent flow systems. As platelets are small fragments produced from megakaryocytes， promoting differentiation efficiency and production of megakaryocytes will optimize platelet generation in vitro. CXCL2 is a chemoattractant chemokine promoting inflammation and angiogenesis， which has been known to be associated with cardiovascular disease and colon cancer， but its function in megakaryocyte differentiation remains unclear. In this study， utilizing a model of differentiation from cord blood-derived CD34⁺ cells to megakaryocytes， CXCL2 expression was decreased by lentivirus-mediated knockdown method， and the differentiation efficiency of megakaryocytes was detected by flow cytometry. Results indicated that early stage of megakaryocyte differentiation was inhibited after CXCL2 knockdown， and this inhibitory effect persisted throughout the differentiation process. This study suggested that CXCL2 plays an important regulatory role in megakaryocyte differentiation， and this finding may exert some implications on megakaryocyte differentiation and platelet generation in vitro.

Mechanism Exploring of Coix Seed Intervention in Fatty Liver Disease Based on Network Pharmacology
Feng DONG, Bingxin HUANGFU, Jia XU, Wentao XU
2023, 13(2): 264-272. DOI: 10.19586/j.2095-2341.2022.0191
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This article aimed to predict the active ingredients and mechanisms of coix seed in the treatment of fatty liver disease， by using network pharmacology and molecular docking technique. The TCMSP database was used to screen the coix seed's active ingredients； PharmMapper， Uniprot， Disgenet， and Venny 2.1.0 databases were used to predict the intersections of relevant gene targets of coix seed and fatty liver disease； String 11.0 was used to create the "protein-protein interaction" network. A cluster-profiler data package was used to perform gene ontology （GO） and the target kyoto encyclopedia of genes and genomes （KEGG） analysis. Cytoscape 3.8.2 software was used to calculate network topology parameters and visualize networks. PubChem， AlphaFold， and CB-DOCK databases were used to obtain small molecular substances and protein structures and perform molecular docking. This research predicted the potential and molecular mechanism of coix seed in the intervention of fatty liver disease. Sitosterol， one of the 9 active ingredients in coix seed， impact fatty liver disease. From the genetic aspect， coix seed interferes with fatty liver disease by regulating genes such as Epidermal Growth Factor Receptor （EGFR）， Caspase-3 （CASP3）， AKT Serine/Threonine Kinase 1 （AKT1）， and Nitric Oxide Synthase 3 （NOS3）； from the pathway aspect，coix seed acts on alcoholic liver disease， endocrine resistance， and estrogen signaling pathways. Coix seed affects fatty liver disease through multiple gene targets and pathways.

Construction of Stable Cell Lines with TOE1 Knockdown and Overexpression and Analysis on its Cellular Localization in Gastric Cancer Cells
Xiaolin SUN, Didi ZHANG, Xinya WANG, Lina ZHANG
2023, 13(2): 273-281. DOI: 10.19586/j.2095-2341.2022.0197
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TOE1 （target of Egr1） is a newly discovered isoenzyme of DEDD superfamily deadenylase Caf1. Construction of its knockdown and overexpression stable cell lines， and analysis of its cellular localization can provide a basis for further research on the function of TOE1 gene. Firstly， the shRNA sequence targeting the silencing of TOE1 gene was inserted into the lentiviral vector pLVX-shRNA2-Puro to construct the lentiviral vector pLVX-shTOE1 that knocks down TOE1 gene. Then， the TOE1 gene sequence was amplified by PCR and cloned into the lentiviral vector pCDH-CMV-MCS-EF1-Puro to construct the lentiviral vector pCDH-TOE1 with overexpression of TOE1 gene. Meanwhile， the eukaryotic expression vector pEGFP-TOE1 was constructed by the same method and transfected into human gastric cancer cell SGC-7901， and the subcellular localization of TOE1 was observed by confocal microscopy. After lentivirus packaging， SGC-7901 cells were infected with pLVX-shTOE1 and pCDH-TOE1 lentivirus， and then SGC-7901-shTOE1 cell line stably silenced TOE1 gene and SGC-7901-oeTOE1 cell line stably overexpressed TOE1 gene were selected by puromycin. Next， the knockdown and overexpression efficiencies of TOE1 at mRNA and protein levels were verified by RT-qPCR and Western blot， respectively. The identification results showed that the expression of TOE1 in SGC-7901-shTOE1 cells was significantly decreased， and the expression of TOE1 in SGC-7901-oeTOE1 cells was obviously increased， suggesting that TOE1-silenced and TOE1-overexpressed SGC-7901 stable gastric cancer cell lines were successfully constructed. Also， the confocal results showed that TOE1 was localized in the nucleus. The results laied a foundation for further exploring the function and mechanism of TOE1 in gastric cancer.

Study on Establishment and Validation of Prognostic Evaluation Model for Hepatocellular Carcinoma Based on Methylomics Database
Guanqun LI, Chaojie LANG, Feng XIA
2023, 13(2): 282-290. DOI: 10.19586/j.2095-2341.2022.0208
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It is a current research hotspot to establish and validate the prognosis related model of hepatocellular carcinoma （HCC） using methylation CG sites. On the R language platform， methylomics data of TCGA-LIHC and GSE136380 were analyzed. Limma package was used to analyze the differential methylation CG sites of TCGA-LIHC and GSE136380， and wgcna package was used to analyze the weighted gene co expression network of GSE136380. The intersection of the three was determined as the key methylation CG site set， and then 83 methylation CG sites were obtained. LASSO-COX risk proportion regression analysis was used to screen six methylation CG sites， including cg15744128， cg24282479， cg17922226， cg13090969， cg23505966 and cg08708079， to construct and validate the prognosis model of HCC after surgical resection. The AUC of the fitting model in training concentration for 1， 3 and 5 years is 0.756 （95% CI：0.644~0.876）， 0.749 （95% CI：0.638~0.854） and 0.797 （95% CI：0.661~0.920） respectively. The AUC of the test set for 1， 3 and 5 years were 0.848 （95% CI： 0.680~0.990）， 0.833 （95% CI：0.697~0.974） and 0.685 （95% CI： 0.553~0.903）， respectively. Kaplan Meier curve showed that it could accurately identify high-risk patients with poor prognosis （logrank test P<0.001） and patients with easy recurrence （logrank test P<0.001） after HCC resection， which is helpful to further develop the treatment plan and benefit the patient.

Screening and Functional Enrichment Analysis of Key Hub Genes in Lynch Syndrome-associated Colorectal Cancer Based on Bioinformatics Analysis
Xingshu ZHU, Junjie HUANG, Caiming WENG, Wangwu LIU, Yongyuan CHEN, Huiyan CHEN, Hu ZHAO, Yu WANG, Chengzhi LIN
2023, 13(2): 291-297. DOI: 10.19586/j.2095-2341.2022.0122
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To screen the differentially expressed genes （DEGs） in Lynch syndrome related colorectal cancer （CRC） by various bioinformatic analysis. The dataset GSE178516 was downloaded from gene expression omnibus （GEO）. DEGs were screened by employing the GEO2R tool， and then intersected with the genes related to CRC and Lynch syndrome from the MalaCards to obtain the common differential genes. Then the functional enrichment analysis and protein-protein interaction （PPI） network was constructed by using String and Cytoscape software. Finally， expressions of the hub genes were verified by easyGEO database. A total of 261 differentially expressed genes were identified， consisting of 117 up-regulated genes and 114 down-regulated genes. After intersection with CRC and Lynch syndrome related genes， a total of 8 genes （TGFBR2， PTEN， POLD1， EXO1， EPCAM， CDKN2B， CCND1， BRCA1） were obtained. GO and KEGG enrichment analysis showed that their functions were mainly enriched in DNA metabolic process regulation， intrinsic apoptosis signaling pathway， mismatch repair， etc. Through PPI analysis， 892 proteins interacting with 8 common DEGs were obtained， including 44 nodes and 630 node interaction relationships. Finally， all 8 genes were statistically differentially expressed in patients with CRC associated with Lynch syndrome. The key Hub genes identified in this study would help us understand the molecular mechanisms behind the occurrence and development of Lynch syndrome combined with CRC， and ultimately provide new candidate targets for the diagnosis and treatment of Lynch syndrome progression to CRC.

The Role of PIM1 Kinase in Cisplatin-induced Acute Kidney Injury
Yuzhen LI, Jiefu ZHU, Xiongfei WU
2023, 13(2): 298-304. DOI: 10.19586/j.2095-2341.2022.0200
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PIM1 is a serine/threonine kinase that participates in the regulation of multiple signal pathways and plays an important role in many diseases. In order to explore the role of PIM1 kinase in cisplatin-induced acute kidney injury， C57BL/6 mice and renal tubular cells were selected as experimental objects. Cisplatin-induced acute kidney injury in mice was established by intraperitoneal injection of cisplatin. The levels of urea nitrogen and creatinine in serum were measured， renal pathology change was assayed by HE staining， qRT-PCR and Western blot were used to detect the expression levels of PIM1 mRNA and PIM1 protein. Renal tubular cells were infected by Adenovirus for overexpression of PIM1， the expression levels of p62、Beclin-1、LC3B were detected by Western blot， CCK-8 and flow cytometry were used to detect the renal tubular cell injury. Results showed that in vivo model， after injection of cisplatin， the levels of serum creatinine， urea nitrogen and tubular injury were significantly increased， the PIM1 mRNA and PIM1 protein expression in kidney tissues were dramatically increased. In vitro model， PIM1 mRNA and PIM1 protein expression levels significantly increased after Adenovirus infection. After cisplatin treated， the cell viability was decreased and the apoptosis was increased， while in the PIM1 overexpression group， the cell viability was increased and the apoptosis was decreased. After cisplatin treated， the expression level of p62 was decreased， the expression levels of Beclin-1， LC3B were increased， and autophagy was activated. Compared with the cisplatin treated group， the expression level of p62 was increased， the expression levels of Beclin-1， LC3B were decreased， and autophagy was weakened in the PIM1 overexpression group. In conclusion， PIM1 was activated in cisplatin-induced acute kidney injury， overexpression of PIM1 alleviated the cisplatin-induced renal tubular cells injury by inhibiting excessive autophagy.

Effects of NRG4 on Proliferation, Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells
Wanling HUANG, Wenqi ZHU, Nini GUO, Nan WANG, Qian REN, Xiaotong MA
2023, 13(2): 305-310. DOI: 10.19586/j.2095-2341.2022.0181
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The aim of the study is to detect the expression of NRG4 in patients with hematopoietic malignancies and malignant hematopoietic cell lines and to investigate its effect on proliferation， apoptosis and clone formation of acute myeloid leukemia （AML） cells．The expression of NRG4 in patients with hematopoietic malignancies and its correlation with the prognosis of patients were analyzed with gene expression omnibus （GEO） and OncoLnc database. Expression of NRG4 mRNA of malignant hematopoietic cell lines were detected by real-time fluorescent quantitative PCR. Lentiviral vector for knockdown of NRG4 were constructed and transduced into AML cell lines Molm-13， KG-1a and Kasumi．The roles of NRG4 in cell proliferation， apoptosis， cell cycle， colony formation were evaluated by cell counting， flow cytometric analysis of Annexin V/7-AAD staining， PI staining， colony-forming assay， respectively. Expression of NRG4 was significantly higher in bone marrow mononuclear cells （BMMCs） from patients with AML compared to those from healthy donors. The survival of AML patients with higher NRG4 expression were significantly shorter than those of AML patients with low NRG4 level（P<0.05）. Knockdown of NRG4 inhibited cell growth and colony formation， promoted apoptosis， and induced cell cycle arrest（P<0.05）. In summary， NRG4 is highly expressed in BMMCs from patients with AML and AML cell lines， and its expression was correlated with poor prognosis in AML patients. NRG4 exerted tumor-promoting roles in AML cells. These findings shed insight into the underlying mechanism of development of AML and provide novel target for AML treatment.

Effects of Melatonin Combined with Cisplatin on Proliferation, Apoptosis and Invasion of Cervical Cancer HeLa Cells
Yunyan SHEN, Qi QIU
2023, 13(2): 311-317. DOI: 10.19586/j.2095-2341.2022.0099
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The aim of this study is to explore the effects of melatonin （MLT） combined with cisplatin （DDP） on the proliferation， apoptosis and invasion of cervical cancer HeLa cells. The cervical cancer HeLa cells were cultured in vitro. The effects of MLT， DDP and their combination on the proliferation of HeLa cells were detected by methyl thiazolyl tetrazolium （MTT） method. HeLa cells were divided into control group， melatonin group， cisplatin group and combination group. The melatonin group was treated with 2.5 mmol·L^-1 melatonin， cisplatin group was treated with 5 μg·mL^-1 cisplatin， and combination group was treated with 2.5 mmol·L^-1 melatonin and 5 μg·mL^-1 cisplatin. The proliferation， apoptosis and invasion of HeLa cells were detected by clone formation assay， flow cytometry， scratch assay and Transwell chamber assay， respectively. The expressions of Ki-67， PCNA， Bcl-2， Bax， N-cadherin， E-cadherin and Vimentin were detected by Western blot. MTT results showed that both MLT and DDP could significantly inhibit the proliferation of HeLa cells （P<0.05）， and the inhibition rate was significantly increased with the increase of their concentration （P<0.05）. Compared with MLT and DDP alone， their combination could significantly increased inhibition rate of HeLa cells proliferation. Compared with control group， clone formation rate， number of invaded cells per unit area， scratch healing rate， and related expression levels of Ki67， PCNA， Bcl-2 and E-cadherin proteins were decreased （P<0.05）， while apoptosis rate and relative expression levels of Bax， N-cadherin and Vimentin proteins were increased in melatonin group and cisplatin group （P<0.05）. Compared with melatonin group and cisplatin group， clone formation rate， number of invaded cells per unit area， scratch healing rate， and relative expression levels of Ki67， PCNA， Bcl-2 and E-cadherin proteins were decreased （P<0.05）， while apoptosis rate and relative expression levels of Bax， N-cadherin and Vimentin proteins were increased in combination group （P<0.05）. Results showed that， both MLT and DDP could inhibit the proliferation and invasion of HeLa cells， and promoted their apoptosis， the effects of their combination were more significant.

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