GTF2H2 Affects the Proliferation and Migration of Hep3B Hepatocellular Carcinoma Cells by Mediating AKT Signal Pathway

doi:10.13523/j.cb.2101026

China Biotechnology

2021, Vol. 41

Issue (6): 4-12 DOI: 10.13523/j.cb.2101026

GTF2H2 Affects the Proliferation and Migration of Hep3B Hepatocellular Carcinoma Cells by Mediating AKT Signal Pathway

OUYANG Qin,LI Yan-meng,XU An-jian,ZHOU Dong-hu,LI Zhen-kun,HUANG Jian(

)

Experimental Center, Beijing Friendship Hospital, Beijing Institute of Clinical Medicine, Beijing 100875, China

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Abstract

Objective: To investigate whether General transcription factor II subunit 2(GTF2H2) affects the proliferation and migration of Hep3B cells and the underlying molecular mechanism. Methods: The GTF2H2 knockdown Hep3B cell model was constructed by transfecting GTF2H2-siRNA. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the GTF2H2 knockdown effect in Hep3B cells. MTS cell proliferation assay kit was used to detect the proliferation ability of GTF2H2 knockdown Hep3B cells. The migration ability of GTF2H2 knockdown Hep3B cells was evaluated by cell Transwell assay. Western blotting was used to detect whether GTF2H2 knockdown affects tumor associated molecular signaling pathway. Results: The proliferation and migration ability of GTF2H2 knockdown Hep3B cells was stronger than that of the controls. Western blotting showed that the expression of p-AKT pathway protein in GTF2H2 knockdown Hep3B cells was significantly increased. Conclusion: GTF2H2 may affect the proliferation and migration ability of Hep3B cells by the regulation of the AKT molecular signal pathway.

Key words： Hep3B GTF2H2 p-AKT Proliferation Migration

Received: 20 January 2021 Published: 06 July 2021

ZTFLH:

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Corresponding Authors: Jian HUANG E-mail: huangj1966@hotmail.com

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	Qin OUYANG
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Cite this article:

OUYANG Qin,LI Yan-meng,XU An-jian,ZHOU Dong-hu,LI Zhen-kun,HUANG Jian. GTF2H2 Affects the Proliferation and Migration of Hep3B Hepatocellular Carcinoma Cells by Mediating AKT Signal Pathway. China Biotechnology, 2021, 41(6): 4-12.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2101026 OR https://manu60.magtech.com.cn/biotech/Y2021/V41/I6/4

Table 1 The primer sequence table of GTF2H2-V1,GTF2H2-V2,GTF2H2-V3 and GAPDH

Fig.1 The GTF2H2 knockdown Hep3B cell model was constructed (a) The expression of GTF2H2 mRNA in control group and GTF2H2 knockdown group was detected by q-RT-PCR (b) The protein expression of GTF2H2 in control group and GTF2H2 knockdown group was detected by Western blotting (c)The statistical graph of Grayscale value of protein strip in control group and GTF2H2 knockdown group^* P<0.05,^** P<0.01

Table 2 Statistical value of GTF2H2 mRNA expression in Hep3B cells treated with GTF2H2-siRNAs

Fig.2 The proliferation ability was enhanced in GTF2H2 knockdown Hep3B cells (a) The proliferation ability of Hep3B cells in the control group and GTF2H2 knockdown group was detected by MTS assay at different time points (b) The protein expression of CyclinD1 and c-Myc in Hep3B cells of control group and GTF2H2 knockdown group was detected by Western blotting assay (c) Statistical results of the protein expression of CylinD1 and c-Myc in the control group and GTF2H2 knockdown group^* P<0.05,^** P<0.01

Table 3 Statistical table of OD value of Hep3B cell proliferation in control group and GTF2H2 knockdown group at different time

Fig.3 The migration ability was promoted in GTF2H2 knockdown Hep3B cells (a) Cell migration assay was used to detect the membrane penetration ability of Hep3B cells in the control group and GTF2H2 knockdown group (b) Comparision of the number of cell migration between the control group and GTF2H2 knockdown group(^* P<0.05,^** P<0.01) (c) Western blotting assay was used to detect the protein expression of MMP9 in Hep3B cells of control group and GTF2H2 knockdown group (d) Statistical results of the protein expression of MMP9 in the control group and GTF2H2 knockdown group(^* P<0.05,^** P<0.01)

Fig.4 The AKT molecular signal pathway was activated in GTF2H2 knockdown Hep3B cells (a) The protein expression of p-AKT,p-ERK,p-P38,p-P65 and p-stat3 in Hep3B cells of control group and GTF2H2 knockdown group was detected by Western blotting assay (b) Statistical results of p-AKT protein expression in the control group and the GTF2H2 knockdown group using total AKT as the standard(^* P<0.05,^** P<0.01) (c) Statistical results of p-ERK protein expression in the control group and GTF2H2 knockdown group using total ERK as the standard(^* P<0.05,^** P<0.01) (d) The statistical results of p-P38,p-P65 and p-stat3 protein expression in the control group and the GTF2H2 knockdown group(^* P<0.05,^** P<0.01)


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