The effect of human papillomavirus 16 E6 gene silenced by small interfering RNA on the expression and the promoter hypermethylation status of E-cadherin in cervical cancer SiHa cell line
10.3760/cma.j.issn.1006-9801.2016.01.002
- VernacularTitle:小干扰 RNA 沉默人乳头瘤病毒16E6基因对子宫颈癌 SiHa 细胞系中 E-钙黏蛋白表达及高甲基化的影响
- Author:
Xian ZHANG
;
Yile CHEN
;
Lesai LI
- Publication Type:Journal Article
- Keywords:
Cadherins glycoprotein;
Human papillomavirus 16E6;
Methylation;
Small interfering RNA;
Cervical neoplasms;
SiHa cell line
- From:
Cancer Research and Clinic
2016;28(1):6-10,20
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the influence of human papillomavirus (HPV) 16 E6 gene silencing by small interfering RNA (siRNA) on the expression and the promoter hypermethylation status of E-cadherin (E-cad) in cervical cancer SiHa cell line. Methods siRNA which used lentivirus as the vector was used to knock down the HPV16E6 gene in cervical cancer SiHa cell line. The expression levels of HPV16E6 mRNA, E-cad mRNA and protein in siRNA-HPV16E6 SiHa cell line were detected by RT-qPCR and Western blot, respectively. Methylation specific PCR (MSP) method was used to detect the methylation status of E-cad gene (CDH1) promoter in siRNA-HPV16E6 SiHa cell line. Results The E-cad mRNA expression levels in siRNA E6 group, empty vector group and blank control group were 4.755±1.085, 1.224± 0.840, 1.327±1.221, respectively. The protein expression levels were 0.616±0.019, 0.325±0.016, 0.299±0.015, respectively. The expressions of E-cad mRNA and protein in siRNA E6 group were significantly higher than those in the empty vector group and blank control group (F = 21.346, P < 0.01; F = 323.398, P < 0.01), and the difference between the empty vector group and blank control group was not statistically significant (P >0.05). After knocking down HPV16E6 gene, the methylation status of E-cad gene was weakly positive, and the intensity of the amplified products was significantly weaker than that in the empty vector group and blank control group, while the unmethylation amplification was positive. Conclusions Knocking down the HPV16E6 gene increases the expression of E-cad in cervical cancer SiHa cell line, and decreases the level of CDH1 promoter methylation. To a certain extent, it partly reverses the hypermethylation status of CDH1 promoter, and causes E-cad to be re-expressed.
