Effect of silencing CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition related protein expressions in HeLa cells of human cervical cancer
10.3760/cma.j.issn.1006-9801.2018.03.002
- VernacularTitle:沉默CXCR4基因对人子宫颈癌HeLa细胞增殖、侵袭及上皮间质转化相关蛋白表达的影响
- Author:
Fengyan LI
1
;
Lijun YU
;
Sanyuan ZHANG
;
Li LI
;
Yan ZHANG
Author Information
1. 山西医科大学第一医院妇科
- Keywords:
Uterine cervical neoplasms;
CXCR4;
Epithelial-mesenchymal transition
- From:
Cancer Research and Clinic
2018;30(3):152-156
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of small interfering RNA (siRNA) CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition factors including Twist, Slug and E-cadherin (E-cad)in HeLa cells of human cervical carcinoma.Methods HeLa cells were cultured in vitro and CXCR4-siRNA was transfected into HeLa cells. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the changes of cell proliferation. Transwell experiment was used to observe the changes of cell invasion, and the expressions of CXCR4,Twist,Slug and E-cad protein were detected by Western blot.Results Compared with the blank control group and the negative control group,CXCR4-siRNA transfected into HeLa cells significantly inhibited the expression of CXCR4 protein (0.959±0.197, 0.932±0.141 vs. 0.379±0.022 respectively; F=16.286, P =0.004). MTT results showed that the proliferation of HeLa cells was inhibited at 48 h and 72 h (48 h: 0.846 ± 0.034, 0.823 ± 0.025 vs. 0.744 ± 0.039, F = 7.379, P= 0.024; 72 h: 0.996±0.026, 0.964± 0.059 vs. 0.829±0.051, F = 10.425, P= 0.011). Transwell results showed that the invasiveness of HeLa cells in CXCR4-siRNA group was inhibited at 48 h and 72 h(cell membrane number,48 h:37.4±8.1,33.6±6.2 vs.25.4 ±3.2, F = 4.830, P= 0.029. 72 h: 48.4 ±7.6, 43.0 ±5.4 vs. 33.4 ±6.7, F = 6.579, P= 0.012). After transfection for 48 h, the expression of Twist and Slug protein was decreased and E-cad protein was increased in CXCR4-siRNA group (Twist: 0.93±0.11, 1.00±0.17 vs. 0.53±0.07, F = 10.962, P= 0.010; Slug: 0.515± 0.084,0.470±0.055 vs.0.190±0.028,F =25.579,P=0.001;E-cad:0.53±0.04,0.55±0.24 vs.1.11±0.14,F =12.494, P= 0.007). Conclusions Silencing CXCR4 gene can inhibit the proliferation and reduce the invasiveness of HeLa cells. Inhibiting EMT of HeLa cells can be achieved by down-regulating the expressions of Twist and Slug protein,and up-regulating the expression of E-cad.
