The mechanism of cisplatin-induced apoptosis in HeLa cells
To study the mechanism of apoptosis induced by cisplatin in vitro in HeLa cells cervical cancer cell line.
The inhibitory effect of cisplatin on HeLa cell growth was analyzed by the MTT assay. Cell apoptosis was measured with flow cytometry and Hoechst 33258 staining following cisplatin treatment. The effect of cisplatin on transcription of HPV E6 was analyzed by RT-PCR and protein expression of E6, P53, p21, Bax and Bcl-2 was studied by Western blots.
Cisplatin inhibited cellular proliferation in a time and dosedependant manner. The sub-G1 peak by flow cytometry showed a higher apoptotic rate in the experimental group compared to the controls and Hoechst 33258 staining indicated that apoptosis was induced by cisplatin. Results of RT-PCR demonstrated that cisplatin decreased transcription of E6. Western Blots showed that cisplatin decreased protein expression of E6 and increased protein expression of P53, p21 and Bax but had no effect on protein expression of Bcl-2.
Cisplatin induces apoptosis and death of HeLa cells through the suppression of HPV E6 and restoration of p53 function.
Keywordscell apoptosis cisplatin flow cytometry HPV E6 Bax Bcl-2 cervix cancer
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