Effect of UbcHlO gene silencing on cell proliferation and cell cycle of lung cancer cell line NCI-H226
10.3724/SP.J.1008.2012.00608
- Author:
Li-ming ZHAO
1
Author Information
1. Department of Respiratory Disease, Changzheng Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
Cell cycle;
Cell proliferation;
Lung neoplasms;
Small interfering RNA;
UbcHlO gene
- From:
Academic Journal of Second Military Medical University
2012;33(6):608-612
- CountryChina
- Language:Chinese
-
Abstract:
Objective To use liposomal siRNA for silencing UbcHlO gene in human lung squamous carcinoma cell line NCI-H226 and to observe the changes of cell proliferation and cell cycle after silencing. Methods Three siRNA sequences targeting different sites of UbcHlO CDS were designed, and the shRNA recombinant plasmids were constructed. The recombinant plasmids were transfected into NCI-H226 cells via lipofectamine2000. UbcHlO mRNA and protein expression was examined by RT-PCR and Western blotting analysis 48 h after transfection, respectively. The viability of NCI-H226 cells was measured using CCK-8 at 24 and 48 h after transfection with the effective siRNA vector, and the cell cycle was detected by flow cytometry 48 h after transfection. Results The recombinant shRNA plasmids were successfully constructed and transfected into NCI-H226 cells. UbcHlO gene mRNA and protein were noticeably decreased in the three groups 48 h after transfection, with the inhibitory effect of No. 2 siRNA sequence (pshRNA2) being the strongest one (86%). The proliferative activity of NCI-H226 cells was significantly decreased 24 and 48 h after the transfection with the pshRNA2 compared with the control group (P<0. 05). Compared with the control group, NCI-H226 cells were blocked in G2 phase 48 h after the transfection with pshRNA2 (P<0. 05). Conclusion UbcHlO gene silencing can significantly inhibit the proliferative activity of NCI-H226 cells and block the cells in G2 phase.
