Effect of expression of high mobility group box-1 inhibited by small hairpin RNA on the invasion and migration of human endometrial carcinoma HEC-1A cells

Objective:To investigate the effect of inhibiting high mobility group box-1(HMGB1) gene expression on the invasion and migration of endometrial carcinoma HEC-1A cells by small hairpin RNA.Methods:Three specific recombinant plasmids of HMGB1(pshRNA-1 /HMGB1,pshRNA-2 /HMGB1,and pshRNA-3/HMGB1) were transfected into the endometrial cancer cell lines HEC-1A by lipofectamine TM 2000.The expression of HMGB1 mRNA and protein was decteted by RTPCR and Western blot.The invasion and migration abilities of transfected HEC-1A cells were evaluated using Transwell assay and wound healing assay.Results:RT-PCR and Western blot revealed that the expression of HMGB1 at both mRNA and protein levels was significantly inhibited by HMGB1-pshRNA targeting sequence 1,2,and 3(P0.05),and the levels of 3 mRNAs in the transfection group were 0.192±0.006,0.055±0.002,and 0.123±0.086,respectively,which were significantly lower than those in the Lipo group(0.268±0.008) and the HMGB1/p-NC group(0.270±0.004).The maximum inhibiton rates of the 3 mRNAs were 28.4%,79.5%,and 54.1%.The levels of 3 HMGB1 proteins in the transfection group were 0.259±0.129,0.032±0.002,and 0.104±0.007,significantly lower than those in the Lipo group(0.347±0.007) and the HMGB1/p-NC group(0.349±0.007),and the maximum inhibitory rates were 25.4%,90.8%,and 70.0%.The transwell chamber assay and wound healing assay showed that the invasion and migration of HEC-1A cells were effectively suppressed by inhibiting HMGB1 expression(P0.05).Conclusion:pshRNA-HMGB1 can effectively inhibit HMGB1 expression at both mRNA and protein levels,and decrease the invasion and migration of endometrial cancer cells.