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.
10.11817/j.issn.1672-7347.2014.01.007
- Author:
Liyuan FENG
1
;
Jiajie WU
Author Information
1. Department of Gynecology and Obstetrics, Xiangya Hospital, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Cell Movement;
Endometrial Neoplasms;
metabolism;
Female;
HMGB1 Protein;
metabolism;
Humans;
Neoplasm Invasiveness;
Plasmids;
RNA, Messenger;
RNA, Small Interfering;
metabolism;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2014;39(1):36-42
- CountryChina
- Language:Chinese
-
Abstract:
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).
CONCLUSION:pshRNA-HMGB1 can effectively inhibit HMGB1 expression at both mRNA and protein levels, and decrease the invasion and migration of endometrial cancer cells.
