Effect and mechanism of TIPE3 interference plasmid on SW480 colorectal canc-er growth
10.3969/j.issn.1000-484X.2017.03.013
- VernacularTitle:TIPE3干扰质粒对SW480结肠癌细胞生长的作用及相关机制探讨
- Author:
Yuhan YE
;
Zhongchen LIU
;
Zhongquan QI
;
Guohong ZHUANG
;
Ping YIN
- Keywords:
TIPE3 shRNA;
Colon cancer;
Apoptosis;
Proliferation
- From:
Chinese Journal of Immunology
2017;33(3):378-383
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effect of interference TIPE3 on the colon cancer cell growth by transfecting SW480 colon cancer cells with the TIPE3 interference plasmid were detected.Methods:Transfecting the constructed TIPE3-shRNA-pSIREN-RetroQ plasmid to SW480 cells.To determine the highest interference efficiency plasmid ,the mRNA and protein levels of recombined plasmid were detected by RT-PCR and Western blot separately and tested the cell proliferation with CCK 8.Meanwhile,apoptosis rate of SW480 cells was determined by flow cytometry assay with AnnexinV-FITC/PI.To further determined the effects of recombined plasmid on cell development ,the level of protein involved in proliferation and apoptosis were detected by Western blot .Results:The most effecient in-terference plasmids were successfully constructed.We found that the cell survival rate decreased when interference TIPE 3 gene express-ing in colorectal cancer cells .Flow cytometry indicated that interefering the expression of TIPE 3 would increase the sensitivity of SW 480 cell to apoptosis induced by aDR5ScFv.The results of Western blot showed that low expression of TIPE 3 would activate caspase3 and downregulate the expression of p-AKT,p-PDK1 and PCNA.Conclusion:Interference TIPE3 could promote apoptosis and inhibit prolif-eration in SW480 colon cancer cells .
