Effect of hypoxia-inducible factor-1α on stemness and epirubicin sensitivity of HepG2 hepatoma cells

Jinjin ZHAO; Haiguang ZHANG; Feifei CUI; Lei WANG; Qingjiang MO; Luyang JIAO

doi:10.3969/j.issn.1001-5256.2021.02.021

Volume 37 Issue 2

Mar. 2021

Turn off MathJax

Article Contents

Article Navigation > Journal of Clinical Hepatology > 2021 > 37(2): 354-357

PDF( 2169 KB)

Effect of hypoxia-inducible factor-1α on stemness and epirubicin sensitivity of HepG2 hepatoma cells

DOI: 10.3969/j.issn.1001-5256.2021.02.021

a.
Department of Clinical Laboratory, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, China
b.
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, China

Received Date: 2020-08-12
Accepted Date: 2020-10-10
Published Date: 2021-02-20

Abstract

Abstract

Objective To investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the stemness and epirubicin sensitivity of hepatoma cells. Methods Hepatoma cells were selected for experiment. HepG2 hepatoma cells transfected with HIF-1α overexpression plasmid were selected as experimental group, and those transfected with pcDNA3.1 empty plasmid were selected as control group; HepG2 cells alone were selected as HepG2 group. Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α; Western blot was used to measure the protein expression of HIF-1α; flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells. The three groups of cells were treated with epirubicin at different concentrations (0, 6.25, 12.5, 25, and 50 μmol/L) for 24 hours; MTT assay was used to measure cell viability, and flow cytometry was used to measure apoptosis after treatment with epirubicin (50 μmol/L). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for further comparison between two groups. Results Compared with the HepG2 group and the control group, the experimental group had a significant increase in the mRNA expression of HIF-1α (both P < 0.001), and Western blot showed high expression of HIF-1α in the experimental group. The percentage of CD133 cells was 0.040%±0.003% in the HepG2 group, 0.030%±0.010% in the control group, and 20.110%±0.600% in the experimental group, and the experimental group had a significantly higher positive rate of CD133⁺ than the HepG2 group and the control group (both P < 0.001). At an epirubicin concentration of 25 and 50 μmol/L, the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group (both P < 0.05). After the treatment with 50 μmol/L epirubicin for 48 hours, the experimental group had a significantly lower cell apoptosis rate than the HepG2 group (67.9%±2.5% vs 93.6%±1.5%, P < 0.001) and the control group (67.9%±2.5% vs 93.0%±1.2%, P < 0.001). Conclusion HepG2 cells are successfully transfected with HIF-1α overexpression plasmid, and HIF-1α can increase the percentage of liver cancer stem cells and improve their resistance to epirubicin.
- Liver Neoplasms, Experimental,
- Hypoxia-Inducible Factor 1, alpha Subunit,
- Epirubicin,
- Stem Cells,
- Drug Resistance, Neoplasm

FullText(HTML)

References(16)

References

[1]	SIA D, VILLANUEVA A, FRIEDMAN SL, et al. Liver cancer cell of origin, molecular class, and effects on patient prognosis[J]. Gastroenterology, 2017, 152(4): 745-761. DOI: 10.1053/j.gastro.2016.11.048
[2]	JIN J. Stem cell treatments[J]. JAMA, 2017, 317(3): 330. DOI: 10.1001/jama.2016.17822
[3]	SUN JH, LUO Q, LIU LL, et al. Liver cancer stem cell markers: Progression and therapeutic implications[J]. World J Gastroenterol, 2016, 22(13): 3547-3557. DOI: 10.3748/wjg.v22.i13.3547
[4]	HUANG Y, LIN D, TANIGUCHI CM. Hypoxia inducible factor (HIF) in the tumor microenvironment: Friend or foe?[J]. Sci China Life Sci, 2017, 60(10): 1114-1124. DOI: 10.1007/s11427-017-9178-y
[5]	QIAN J, RANKIN EB. Hypoxia-induced phenotypes that mediate tumor heterogeneity[J]. Adv Exp Med Biol, 2019, 1136: 43-55.
[6]	SAMANTA D, PARK Y, NI X, et al. Chemotherapy induces enrichment of CD47⁺/CD73⁺/PDL1⁺ immune evasive triple-negative breast cancer cells[J]. Proc Natl Acad Sci U S A, 2018, 115(6): e1239-e1248. DOI: 10.1073/pnas.1718197115
[7]	LU H, SAMANTA D, XIANG L, et al. Chemotherapy triggers HIF-1-dependent glutathione synthesis and copper chelation that induces the breast cancer stem cell phenotype[J]. Proc Natl Acad Sci U S A, 2015, 112(33): e4600-e4609. DOI: 10.1073/pnas.1513433112
[8]	LAN J, LU H, SAMANTA D, et al. Hypoxia-inducible factor 1-dependent expression of adenosine receptor 2B promotes breast cancer stem cell enrichment[J]. Proc Natl Acad Sci U S A, 2018, 115(41): e9640-e9648. DOI: 10.1073/pnas.1809695115
[9]	SEMENZA GL. The genomics and genetics of oxygen homeostasis[J]. Annu Rev Genomics Hum Genet, 2020, 21: 183-204. DOI: 10.1146/annurev-genom-111119-073356
[10]	MARIE-EGYPTIENNE DT, LOHSE I, HILL RP. Cancer stem cells, the epithelial to mesenchymal transition (EMT) and radioresistance: Potential role of hypoxia[J]. Cancer Lett, 2013, 341(1): 63-72. DOI: 10.1016/j.canlet.2012.11.019
[11]	NOZAWA-SUZUKI N, NAGASAWA H, OHNISHI K, et al. The inhibitory effect of hypoxic cytotoxin on the expansion of cancer stem cells in ovarian cancer[J]. Biochem Biophys Res Commun, 2015, 457(4): 706-711. DOI: 10.1016/j.bbrc.2015.01.053
[12]	BEN-PORATH I, THOMSON MW, CAREY VJ, et al. An embryonic stem cell-like gene expression signature in poorly differentiated aggressive human tumors[J]. Nat Genet, 2008, 40(5): 499-507. DOI: 10.1038/ng.127
[13]	ALDERSON D, CUNNINGHAM D, NANKIVELL M, et al. Neoadjuvant cisplatin and fluorouracil versus epirubicin, cisplatin, and capecitabine followed by resection in patients with oesophageal adenocarcinoma (UK MRC OE05): An open-label, randomised phase 3 trial[J]. Lancet Oncol, 2017, 18(9): 1249-1260. DOI: 10.1016/S1470-2045(17)30447-3
[14]	SONG B, BIAN Q, SHAO CH, et al. Ulinastatin reduces the resistance of liver cancer cells to epirubicin by inhibiting autophagy[J]. PLoS One, 2015, 10(3): e0120694. DOI: 10.1371/journal.pone.0120694
[15]	LI J, DUAN B, GUO Y, et al. Baicalein sensitizes hepatocellular carcinoma cells to 5-FU and Epirubicin by activating apoptosis and ameliorating P-glycoprotein activity[J]. Biomed Pharmacother, 2018, 98: 806-812. DOI: 10.1016/j.biopha.2018.01.002
[16]	PLAKS V, KONG N, WERB Z. The cancer stem cell niche: How essential is the niche in regulating stemness of tumor cells?[J]. Cell Stem Cell, 2015, 16(3): 225-238. DOI: 10.1016/j.stem.2015.02.015

Relative Articles

Supplements(0)

Cited By

Proportional views

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Figures(3) / Tables(1)

Get Citation

PDF

XML

Article Metrics

Article views (472) PDF downloads(30)

Effect of hypoxia-inducible factor-1α on stemness and epirubicin sensitivity of HepG2 hepatoma cells

DOI: 10.3969/j.issn.1001-5256.2021.02.021

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Effect of hypoxia-inducible factor-1α on stemness and epirubicin sensitivity of HepG2 hepatoma cells

DOI: 10.3969/j.issn.1001-5256.2021.02.021

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

About Journal

Submission Guideline

Journal Online

Hepatobiliary College

Guidelines

Export File

Citation

Format

Content