Effect Of Expression Of High Mobility Group Box-1Inhibited 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 lipofectamineTM2000. The expression of HMGB1mRNA and protein were decteted by RT-PCR and Western-blot.The invasion and migration abilities of transfected HEC-1A cells were evaluated using Transwell assay and wound healing assay.Results:1. RT-PCR revealed that the expression of HMGB1at mRNA level was significantly inhibited by HMGB1-pshRNA targeting sequence1,2, and3(P<0.05), and the levels of3mRNAs in the transfection group were0.192±0.006,0.055±0.002, and0.123±0.004, 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 inhibition rates of the3mRNAs were28.9%,79.6%, and54.4%.2.The levels of3HMGB1proteins in the transfection group were0.259±0.013,0.032±0.002, and0.104±0.007, significantly lower than those in the Lipo group (0.347±0.007) and the HMGBl/p-NC group (0.349±0.007), and the maximum inhibitory rates were25.8%,90.8%, and70.2%.3.The transwell chamber assay showed that the invasion and migration of HEC-1A cells was effectively suppressed by inhibiting HMGB1expression (P<0.05). The average migration cell count of pshRNA-2/HMGB1infected cells was (20±1), much less than of the Lipo group (36±1) and the HMGB1/p-NC group (36±2). No significant difference was found between Lipo group and HMGB1/p-NC group (P>0.05).4.The wound healing assay showed that the migration of HEC-1A cells was effectively suppressed by inhibiting HMGB1expression (P<0.05). The average migration rate of pshRNA-2/HMGB1infected cells was (25.75%±1.70%), much lower than of the Lipo group (65.25%±1.70%) and the HMGB1/p-NC group (64.75%±4.57%). No significant difference was found between Lipo group and HMGB1/p-NC group (P>0.05). Conclusions:1.pshRNA-HMGB1can effectively inhibit HMGB1expression at both mRNA and protein levels, especially targeting sequence2.2. The invasion and migration of endometrial cancer cells was effectively decreased by pshRNA-2/HMGB1infected cells, which indicated that HMGB1maybe play an important part in the occurrence and development of endometrial cancer by incontrollable of the cell invasion and migration.