[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 (P<0.05), 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 (P<0.05).


CONCLUSIONS
pshRNA-HMGB1 can effectively inhibit HMGB1 expression at both mRNA and protein levels, and decrease the invasion and migration of endometrial cancer cells.
Department of Gynecology and Obstetrics, Xiangya Hospital, Central South University, Changsha 410008, China.
Liyuan
L
Feng
Jiajie
J
Wu
D045744:Cell Line, Tumor; D002465:Cell Movement; D016889:Endometrial Neoplasms / Q000378:metabolism*; D005260:Female; D024243:HMGB1 Protein / Q000378:metabolism*; D006801:Humans; D009361:Neoplasm Invasiveness; D010957:Plasmids; D012333:RNA, Messenger; D034741:RNA, Small Interfering / Q000378:metabolism*; D014162:Transfection
D024243:HMGB1 Protein; C585491:HMGB1 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering