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Published on: April 29, 2014
Study on the effects of allitride and its mechanisms on renal cell carcinoma in vitro
Hui Liu1, Rong-pei Wu, Xiao-fei Li
1Department of Urology, Henan University Huaihe hospital, Kaifeng, 475000, China.
Objective:
To study the killing effect of allitride on human renal cell carcinoma cell line Ketr-3 and its possible mechanisms.
Methods:
The Ketr-3 cells were treated with allitride and the morphological changes were observed with inverted microscope. The cytotoxicity was estimated through theamine blue tetrazolium bromide (MTT). Apoptotic cells were detected by in situ cell apoptosis detection kit, and confirmed by flow cytometry. Changes of apoptosis rate cell cycle were assessed by flow cytometry. Caspase-3 (cysteineaspartate specific proteinase) mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and Caspase-3 protease activity was estimated with colorimetry.
Results:
MTT assay and morphological changes confirmed the killing effect of allitride on Ketr-3 cellline. FCM also showed that S-phase and G2/M-phase arrest was induced. RT-PCR and colorimetry confirmed that there was apparently a rise of Caspase-3 mRNA and Caspase-3 protease activity.
Conclusion:
Allitride could kill Ketr-3 effectively by inducing apoptosis. Cell cycle arrest and up-regulation of Caspase-3 may play an important role in the mechanisms of killing effect.
