HPV16 E6调控miR-23a表达促进宫颈癌细胞侵袭、迁移

doi:10.3969/j.issn.1006-5725.2024.02.004

Abstract

Abstract:

Objective To investigate the effects of high-risk human papillomavirus 16 E6 protein （HPV16 E6 protein） on invasion and migration of cervical cancer SiHa cells via regulating the expression of expression miR-23a. Methods Tissue samples from 100 patients with cervical cancer HPV-negative， 100 HPV-positive patients， and 100 paracancerous normal tissues were collected； cervical cancer SiHa cells were divided into blank group， E6 overexpression group， negative transfection group， and E6 + miR-23a mimics group. The expression of miR-23a and HPV16 E6 mRNA were detected by qRT-PCR； MTT assay was used to detect the cell proliferation inhibition rate； flow cytometry to detect the apoptosis； Transwell chamber assay to detect cell invasion， and scratch test to detect the ability of cell migration. The expression of HPV16 E6， apoptosis related proteins （Caspase-3， Bax， Bcl-2）， and migration related proteins （MMP-2， MMP-9） was detected by WB. Results The expression level of miR-23a was decreased in cervical cancer tissues， and that was lower in HPV positive cervical cancer tissues. Overexpression of E6 decreased the expression level of miR-23a， cell proliferation inhibition rate， apoptosis rate， Caspase-3 and Bax protein expression， and increased the expression of Bcl-2 protein， scratch healing rate， invasion cell number， MMP-2， MMP-9 protein expression （P < 0.05）； miR-23a mimics reversed the effects of E6 overexpression on the above indicators. Conclusion HPV16 E6 promotes the invasion and migration of cervical cancer cells， which may be related to the regulation of miR-23a expression.

Key words: human papillomavirus 16 E6 protein, miR-23a, cervical cancer, invasion, migration

CLC Number:

R737.33

Min WANG,Dan MU,Dejun KONG,Li YANG,Lu YE,Dan HE. HPV16 E6 regulates miR-23a expression and promotes invasion and migration of cervical cancer cells[J]. The Journal of Practical Medicine, 2024, 40(2): 146-152.

Figures/Tables 9

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References 26

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组别	HPV16 E6/ β-actin	HPV16 E6/GAPDH	miR-23a/U6
空白组	1.02 ± 0.20	1.01 ± 0.20	1.03 ± 0.21
阴性转染组	1.01 ± 0.19	1.02 ± 0.21	1.03 ± 0.18
E6过表达组	1.56 ± 0.25^ab	1.52 ± 0.24^ab	0.32 ± 0.06^ab
E6+miR-23a mimics组	1.55 ± 0.26^ab	1.54 ± 0.25^ab	0.65 ± 0.13^abc
F值	5.659	5.200	14.494
P值	0.022	0.028	0.001

组别	细胞凋亡率（%）	凋亡相关因子蛋白表达
组别	细胞凋亡率（%）	Caspase-3/GAPDH	Bax/GAPDH	Bcl-2/GAPDH
空白组	18.28 ± 1.23	0.98 ± 0.15	1.03 ± 0.14	0.94 ± 0.07
阴性转染组	19.01 ± 1.25	1.01 ± 0.10	0.99 ± 0.13	1.00 ± 0.08
E6过表达组	8.44 ± 0.75^ab	0.26 ± 0.07^ab	0.30 ± 0.05^ab	1.64 ± 0.11^ab
E6+miR-23a mimics组	15.33 ± 1.01^abc	0.71 ± 0.08^abc	0.65 ± 0.08^abc	1.25 ± 0.09^abc
F值	59.854	33.041	30.685	38.636
P值	0.001	0.000	0.000	0.000

组别	侵袭细胞数（个）	划痕愈合率（%）	细胞侵袭相关蛋白表达
组别	侵袭细胞数（个）	划痕愈合率（%）	MMP-2/GAPDH	MMP-9/GAPDH
空白组	85.40 ± 4.50	19.86 ± 2.11	1.02 ± 0.13	0.96 ± 0.12
阴性转染组	83.70 ± 4.73	18.81 ± 2.01	0.96 ± 0.10	0.93 ± 0.09
E6过表达组	120.94 ± 6.55^ab	31.35 ± 3.07^ab	1.65 ± 0.18^ab	1.61 ± 0.16^ab
E6+miR-23a mimics组	102.66 ± 5.03^abc	25.58 ± 1.03^abc	1.30 ± 0.11^abc	1.29 ± 0.10^abc
F值	32.883	21.194	16.710	21.114
P值	0.000	0.000	0.001	0.000

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HPV16 E6 regulates miR-23a expression and promotes invasion and migration of cervical cancer cells

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