Mechanistic Study on Total Flavonoids from Hedyotis diffusa Willd. Inhibiting Lung Cancer Through the CDCA8-p53-glycolysis Axis

doi:10.19586/j.2095-2341.2025.0147

Current Biotechnology ›› 2026, Vol. 16 ›› Issue (4): 979-989.DOI: 10.19586/j.2095-2341.2025.0147

• Articles • Previous Articles

Mechanistic Study on Total Flavonoids from Hedyotis diffusa Willd. Inhibiting Lung Cancer Through the CDCA8-p53-glycolysis Axis

Yue LUO¹(), Xing JIANG¹, Chao WANG¹, Yingqiu WU¹, Hongwei LUO¹, Lu ZHANG²()

^1.Mianzhu People's Hospital，Sichuan Deyang 618000，China
^2.Chongqing College of Traditional Chinese Medicine，Chongqing 402760，China

Received:2025-10-27 Accepted:2026-03-10 Online:2026-07-25 Published:2026-09-11
Contact: Lu ZHANG

Abstract

Abstract:

This study aimed to investigate the molecular mechanism by which total flavonoids of Hedyotis diffusa Willd. （HDF） exert anti-lung cancer effects through modulation of the CDCA8-p53-glycolysis axis. A549 cell models with CDCA8 knockdown/overexpression were constructed， and cell proliferation， colony formation， migration， and invasion abilities were assessed. Quantitative real-time PCR（qPCR） and Western blot were employed to detect the expression of CDCA8， p53， and key glycolytic enzymes. Glucose uptake， lactate production， and ATP levels were measured using colorimetric assays. Cells were treated with varying concentrations of HDF， and their effects on the CDCA8-p53-glycolysis axis and malignant cell phenotypes were evaluated. The results showed that CDCA8 knockdown significantly inhibited A549 cell proliferation， migration， and invasion （P<0.01）， upregulated p53 and p-p53 （Ser15） protein expression （P<0.05）， downregulated the expression of key glycolytic enzymes GLUT1， HK2， and PKM2 （P<0.05）， and reduced glucose uptake， lactate production， and ATP levels （P<0.05）； CDCA8 overexpression exhibited opposite effects. The IC₅₀ of HDF against A549 cells was 1.6 mg·mL^-1. HDF dose-dependently downregulated CDCA8， upregulated p53/p-p53， inhibited the expression and metabolism of key glycolytic enzymes （P<0.05）， and significantly suppressed cell proliferation， migration， and invasion abilities （P<0.05）. The results indicated that HDF exerts anti-lung cancer effects by downregulating CDCA8 expression， activating the p53 signaling pathway， and inhibiting glycolysis.

Key words: total flavonoids of Hedyotis diffusa Willd., lung cancer, CDCA8, p53, glycolysis, A549 cells

CLC Number:

Q257

Yue LUO, Xing JIANG, Chao WANG, Yingqiu WU, Hongwei LUO, Lu ZHANG. Mechanistic Study on Total Flavonoids from Hedyotis diffusa Willd. Inhibiting Lung Cancer Through the CDCA8-p53-glycolysis Axis[J]. Current Biotechnology, 2026, 16(4): 979-989.

Add to citation manager EndNote|Ris|BibTeX

URL: https://swjsjz.magtechjournal.com/EN/10.19586/j.2095-2341.2025.0147

https://swjsjz.magtechjournal.com/EN/Y2026/V16/I4/979

Figures/Tables 15

Table 1 Primer sequences

基因	引物序列（5'→3'）
CDCA8	F: 5'-TCCTCGTCCCTACCCAGTTT-3' R: 5'-GCCTCCGTAAGGAGTTGGTC-3'
p53	F: 5'-GCGCACAGAGGAAGAGAATC-3' R: 5'-GGCCAACTTGTTCAGTGGAG-3'
HK2	F: 5'-GGGAGGTCTCCACTGAGGAT-3' R: 5'-ACGTACAACTGACCATTCATTACA-3'
PKM2	F: 5'-CCTGATAGCTCGTGAGGCTG-3' R: 5'-AGGCTCGCACAAGTTCTTCA-3'
GLUT1	F: 5'-AATGGTTCCATGCCTGAGGG-3' R: 5'-AAGGCAAGTGTCTCGACAGG-3'
β-actin	F: 5'-GGACTTCGAGCAAGAGATGG-3' R: 5'-ACATCTGCTGGAAGGTGGAC-3'

Fig. 1 Validation of CDCA8 knockdown and over-expression in A549 cells

Fig. 2 Effect of CDCA8 on the p53 signaling pathway

Fig. 3 Effect of CDCA8 on glycolysis in A549 cells

Fig. 4 Effect of CDCA8 knockdown and overexpression on A549 cell proliferation activity assessed by the CCK-8 assay

Fig. 5 Colony formation assay

Fig. 6 A549 detection of cell migration and invasion abilities

Fig. 7 Determination of total flavonoid concentration in Hedyotis diffusa Willd.

Fig. 8 Effect of total flavonoids from Hedyotis diffusa Willd. (HDF) treatment for 24 h on A549 cell proliferation assessed by the CCK-8 assay

Fig. 9 Effect of total flavonoids from Hedyotis diffusa Willd. on CDCA8 expression in A549 cells

Fig. 10 Effect of total flavonoids from Hedyotis diffusa Willd. on the p53 signaling pathway in A549 cells

Fig. 11 Effect of total flavonoids from Hedyotis diffusa Willd. on glycolysis in A549 cells

Fig. 12 Effect of different concentrations of total flavonoids from Hedyotis diffusa Willd. on A549 cell proliferation assessed by the CCK-8 assay

Fig. 13 Effect of different concentrations of HDF on colony formation

Fig. 14 Effect of different concentrations of HDF on cell migration and invasion abilities

References 26

[1]	OLIVER A L. Lung cancer: epidemiology and screening[J]. Surg. Clin. North Am., 2022, 102(3): 335-344.
[2]	罗添乐,罗斌,田建辉.中医药防治肺癌的理论研究进展[J].中医肿瘤学杂志,2019,1(2):1-6.
	LUO T L, LUO B, TIAN J H. Advances in theoretical research on prevention and treatment of lung cancer with traditional Chinese medicine[J]. J. Oncol. Chin. Med., 2019, 1(2): 1-6.
[3]	陈浩方,雷旭东,党会芬,等.中医专方治疗肺癌用药规律研究[J].甘肃医药,2022,41(2):134-136.
	CHEN H F, LEI X D, DANG H F, et al.. The study of the law of drug use for the treatment of lung cancer by Chinese medicine[J]. Gansu Med. J., 2022, 41(2): 134-136.
[4]	曾珠,何成诗,孙剑峰,等.白花蛇舌草对A549肺癌干细胞增殖、凋亡及细胞周期的影响[J].重庆医学,2019,48(13):2202-2205.
	ZENG Z, HE C S, SUN J F, et al.. Effect of Hedyotis diffusa on proliferation, apoptosis and cell cycle of A549 lung cancer stem cells[J]. Chongqing Med., 2019, 48(13): 2202-2205.
[5]	LIN J, LI Q, CHEN H, et al.. Hedyotis diffusa Willd. extract suppresses proliferation and induces apoptosis via IL-6-inducible STAT3 pathway inactivation in human colorectal cancer cells[J]. Oncol. Lett., 2015, 9(4): 1962-1970.
[6]	LI H, LAI Z, YANG H, et al.. Hedyotis diffusa Willd. inhibits VEGF-C-mediated lymphangiogenesis in colorectal cancer via multiple signaling pathways[J]. Oncol. Rep., 2019, 42(3): 1225-1236.
[7]	高艳艳,王贵平,李春玲,等.白花蛇舌草提取物对肉鸡淋巴细胞增殖功能的影响[J].动物医学进展,2010,31(10):43-46.
	GAO Y Y, WANG G P, LI C L, et al.. Effect of Oldenlandia extract on lymphocyte proliferation in chickens[J]. Prog. Vet. Med., 2010, 31(10): 43-46.
[8]	王雪,夏顺利,刘景楠,等.白花蛇舌草总黄酮大孔树脂纯化工艺及其体内抑瘤作用的研究[J].中医药信息,2021,38(3):16-21.
	WANG X, XIA S L, LIU J N, et al.. Study on purification process of total flavonoids from Oldenlandia diffusa by macroporous resin and its tumor inhibition in vivo [J]. Inf. Tradit. Chin. Med., 2021, 38(3): 16-21.
[9]	PENG B, ZHANG J, XIANG Y. CDCA8 and its multifaceted role in tumorigenesis[J/OL]. Biomed. Pharmacother., 2025, 185: 117951[2025-12-07]. .
[10]	GU H, GAO X, HAN W, et al.. Overexpression of CDCA8 predicts poor prognosis and drug insensitivity in lung adenocarcinoma[J/OL]. BMC Med. Genom., 2024, 17(1): 265[2025-12-07]. .
[11]	ZHOU Q, HUANG W, XIONG J, et al.. CDCA8 promotes bladder cancer survival by stabilizing HIF1α expression under hypoxia[J/OL]. Cell Death Dis., 2023, 14: 658[2025-12-07]. .
[12]	XIANG C, SUN W H, KE Y, et al.. CDCA8 contributes to the development and progression of thyroid cancer through regulating CDK1[J]. J. Cancer, 2022, 13(7): 2322-2335.
[13]	罗悦,雒洪伟,姜星,等.基于转录组学探讨白花蛇舌草总黄酮抗肺癌A549细胞生长机制[J].药学研究,2023,42(11):857-864.
	LUO Y, LUO H W, JIANG X, et al.. Transcriptomic study of total flavonoids of Hedyotis diffusa Willd. against the growth of lung cancer cells[J]. J. Pharm. Res., 2023, 42(11): 857-864.
[14]	LEVINE A J. p53: 800 million years of evolution and 40 years of discovery[J]. Nat. Rev. Cancer, 2020, 20(8): 471-480.
[15]	LIU J, ZHANG C, HU W, et al.. Tumor suppressor p53 and metabolism[J]. J. Mol. Cell Biol., 2019, 11(4): 284-292.
[16]	LI X, WU L, ZOPP M, et al.. p53-TP53-induced glycolysis regulator mediated glycolytic suppression attenuates DNA damage and genomic instability in Fanconi anemia hematopoietic stem cells[J]. Stem Cells, 2019, 37(7): 937-947.
[17]	ZHANG C, LIU J, WU R, et al.. Tumor suppressor p53 negatively regulates glycolysis stimulated by hypoxia through its target RRAD[J]. Oncotarget, 2014, 5(14): 5535-5546.
[18]	KIM J, YU L, CHEN W, et al.. Willd-type p53 promotes cancer metabolic switch by inducing PUMA-dependent suppression of oxidative phosphorylation[J]. Cancer Cell, 2019, 35(2): 191-203.
[19]	LIU Y, GU W. The complexity of p53-mediated metabolic regulation in tumor suppression[J]. Semin. Cancer Biol., 2022, 85: 4-32.
[20]	HERNÁNDEZ-RESÉNDIZ I, GALLARDO-PÉREZ J C, LÓPEZ-MACAY A, et al.. Mutant p53R248Q downregulates oxidative phosphorylation and upregulates glycolysis under normoxia and hypoxia in human cervix cancer cells[J]. J. Cell. Physiol., 2019, 234(5): 5524-5536.
[21]	HANAHAN D, WEINBERG R A. Hallmarks of cancer: the next generation[J]. Cell, 2011, 144(5): 646-674.
[22]	VANDER HEIDEN M G, CANTLEY L C, THOMPSON C B. Understanding the Warburg effect: the metabolic requirements of cell proliferation[J]. Science, 2009, 324(5930): 1029-1033.
[23]	王彩虹,陈剑飞,鞠久君,等.PKM2促进肿瘤发展的作用机制研究[J].生物技术进展,2022,12(3):411-418.
	WANG C H, CHEN J F, JU J J, et al.. The mechanisms of PKM2 promoting tumor progression[J]. Curr. Biotechnol., 2022, 12(3): 411-418.
[24]	HUANG F, PANG J, XU L, et al.. Hedyotis diffusa injection induces ferroptosis via the Ba _x /Bcl2/VDAC2/3 axis in lung adenocarcinoma[J/OL]. Phytomedicine, 2022, 104: 154319[2025-12-07]. .
[25]	李佳林.白花蛇舌草总黄酮联合顺铂对肺癌A549细胞增殖、凋亡的影响及可能机制[D].衡阳:南华大学,2018.
[26]	孙晓春,郭佳宸,孙琛,等.桔梗、白花蛇舌草不同组分对人肺癌细胞A549的抑制作用[J].中国农学通报,2019,35(1):144-148.
	SUN X C, GUOJIA C, SUN C, et al.. The inhibitory effect of extracts from Platycodon grandiflorum and Hedyotis diffusa willd on human lung cancer cells A549[J]. Chin. Agric. Sci. Bull., 2019, 35(1): 144-148.

Mechanistic Study on Total Flavonoids from Hedyotis diffusa Willd. Inhibiting Lung Cancer Through the CDCA8-p53-glycolysis Axis

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 15

References 26

Related Articles 7

Recommended Articles

Metrics

Comments

[1]	Hongfei LIU, Wen ZHAO, Huiying YANG. ALKBH5 Affects Cisplatin Chemosensitivity in Non-small Cell Lung Cancer by Regulating CPNE1 m⁶A Methylation [J]. Current Biotechnology, 2026, 16(2): 412-421.
[2]	Yanjie WANG, Bo QIU. The Role and Research Progress of Gene P53 in Osteosarcoma [J]. Current Biotechnology, 2025, 15(2): 241-246.
[3]	Xuan ZHAO, Limei REN, Xiaoru WANG, Guangxin HAN, Dandan GU, Yasen YAO, Yonghao QI. Effects of siRNA Targeting to Interfere with the Expression of TRAF6 on the Proliferation and Apoptosis of Lung Cancer Cells [J]. Current Biotechnology, 2024, 14(5): 875-881.
[4]	Qianting YANG, Xiaoying TIAN, Junfang ZHANG, Bingshe HAN. Effect of tp53 Knockout on High Temperature Tolerance and Swimming Capacity in Zebrafish [J]. Current Biotechnology, 2023, 13(4): 580-587.
[5]	Jian DU, Zhigang TIAN, Hua LIN, Keliang XIE. Effect of Hydrogen Inhalation on Postoperative Delirium in Patients Undergoing Radical Surgery for Lung Cancer [J]. Current Biotechnology, 2022, 12(4): 510-515.
[6]	LI Fangfang, MU Dengcai,YANG Chunyan, WANG Lei, SHEN Hao, ZHENG Shangyong^*. Expression and Function of KIF26B in the Development of Non-small Cell Lung Cancer [J]. Curr. Biotech., 2020, 10(3): 273-283.
[7]	ZHANG Haitao, ZHANG Yuxin*. miR-148a Inhibits Glycolysis and Cell Proliferation of Breast Cancer Cells Through Targeted Regulation of Hexokinase 2 Gene [J]. Curr. Biotech., 2020, 10(2): 176-184.