Lentivirus mediated RNA interference inhibits mesothelin expression in ovarian cancer cells and cell proliferation
- VernacularTitle:慢病毒介导的RNAi对卵巢癌细胞增殖及间皮素表达的影响
- Author:
Li WANG
;
Na LI
;
Xiaohua WU
- Publication Type:Journal Article
- Keywords:
mesothelin;
RNA interference;
lentivirus;
ovarian neoplasms;
cell proliferation
- From:
Chinese Journal of Cancer Biotherapy
1995;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a recombinant lentivirus plasmid of RNA interference targeting (MSLN) gene and to observe its effect on MSLN expression in human ovarian cancer cell line OVCAR-3 and its effect on cell proliferation. Methods: According to the Genbank information of MSLN, four RNA interfering sequences and a negative sequence were designed and inserted into plasmid pRNAT-U6.2/Lenti and 5 kinds of plasmids were packaged: LV-MSLN-negative,LV-MSLN-shRNA1, LV-MSLN-shRNA2, LV-MSLN-shRNA3, and LV-MSLN-shRNA4; and they were used to transfect OVCAR-3 cells. Western blotting and indirect immunofluorescence were then used to investigate the interfering efficiency. The plasmid with high interfering efficiency was packaged. The cell proliferation test and clone-forming test was used to assess the changes in cell proliferation. Results: DNA sequencing showed that the sequences of 5 recombinant lentivirus plasmids were correct. Lentivirus packaging was successfully done. Western blotting analysis confirmed that LV-MSLN-shRNA4 had the highest interfering efficiency (90%). MSLN specifically bound to cytomembrane of OVCAR-3 cells. Expression of MSLN in the interfered cells (OVC-shRNA) was weaker than that in the control cells (OVC-neg,OVC). OVC-shRNA cells([11.2?1.3]?105) grew slowly compared to OVC-neg cells([20.5?2.5]?105) and OVC cells([21.9?2.3]?105) (P
