Effect of silencing CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition related protein expressions in HeLa cells of human cervical cancer / 肿瘤研究与临床

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 h37.4±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-cad0.53±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.