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Objective:The aim of the study was to explore the activities of cis9,trans11-CLA(C9,t11-CLA) and trans10,cis12-CLA(t10,c12-CLA) inhibiting tumor,and investigate their relationships with PPARγ and apoptotic proteins,and mechanism of anti-cancer.Methods:The inhibitory rate,cell growth curve and apoptotic morphological observation of MCF-7 cells were obtained by MTT assay,trypan blue staining and Hoechst33342 fluorescence staining.The apoptotic rate and cell cycle were detected with flow cytometry.Transcriptional level of genes was detected with RT-PCR semi-quantitative method,and Western blot was performed to detect proteins levels.Results:The two CLA isomers could reduce cell proliferation(P < 0.05),increase apoptotic rate(P < 0.05),and increase obviously the transcriptional and protein levels of PPARγ(P < 0.01).The synchronism and correlation between the effects of CLA to PPARγ and apoptotic proteins Bax,Bcl-2,Caspase 3 changes were found with the dose-and time-dependent manners.There was cooperative relation between the levels of PPARγ and the rates of Bax/Bcl-2,Caspase 3(small fragment) by experiments of PPARγ inhibitor GW9662 and ligand Rosiglitazone.Conclusion:The apoptotic pathway of PPARγ-Bcl-2-Caspase 3 signaling was found.The C9,t11-CLA and t10,c12-CLA could inhibit MCF-7 cell proliferation and promote apoptosis via activating PPARγ-Bcl-2-Caspase 3 pathway.CLA may be a kind of activator of PPARγ.
Objective: The aim of the study was to explore the activities of cis9, trans11-CLA (C9, t11-CLA) and trans10, cis12- CLA (t10, c12- CLA) inhibiting tumor, and investigate their relationships with PPARγ and apoptotic proteins , and mechanism of anti-cancer. Methods: The inhibitory rate, cell growth curve and apoptotic morphological observation of MCF-7 cells were obtained by MTT assay, trypan blue staining and Hoechst 33342 fluorescence staining. apoptotic rate and cell cycle were detected with flow cytometry. Transcription level of genes was detected with RT-PCR semi-quantitative method, and Western blot was performed to detect protein levels. Results: The two CLA isomers could reduce cell proliferation (P <0.05), increase apoptotic rate (P <0.05 ), and increase obviously the transcriptional and protein levels of PPARγ (P <0.01). The synchronism and correlation between the effects of CLA to PPARγ and apoptotic proteins Bax, Bcl-2, Caspase 3 changes were found with the dose- and time- dependent manners.The re was the relationship between the levels of PPARγ and the rates of Bax / Bcl-2, Caspase 3 (small fragment) by experiments of PPARγ inhibitor GW9662 and ligand Rosiglitazone. Confluence: The apoptotic pathway of PPARγ-Bcl-2-Caspase 3 signaling was found. C9, t11-CLA and t10, c12-CLA could inhibit a MCF-7 cell proliferation and promote apoptosis by activating PPARγ-Bcl-2-Caspase 3 pathway. CLA may be a kind of activator of PPARγ.