The Relationship Between The Changes Of The Alternative Splicing Of The MRNAs Of CDC25B,C And The Resistance To Cisplatin In Cervical Cancer Cells

Cervical cancer is the most common malignant tumor in women, seriously threatenning the health and life of women. Cisplatin is commonly used in the treatment of cervical cancer as a kind of chemotherapeutic drug, but the resistance of the cancer cells to cisplatin greatly reduces the treatment effect. With cisplatin in the treatment of cervical carcinoma SiHa and C33a cell lines, the alternative splicing of the mRNA of CDC25B and CDC25C genes are changed. CDC25B and CDC25C play an important role in regulating cell cycle checkpoint function under normal physiological conditions and in response to DNA damage. Therefore, it is inferred that the alternative splicing changes of the mRNA of CDC25B and CDC25C genes are likely to be related to cell tolerance to cisplatin. This research intends to explore whether the main splicing isoforms of the CDC25B and CDC25C have anti apoptosis or apoptosis. Whether these splicing changes increase the resistance of the cervical cancer cells to cisplatin. The results and information obtained will help to clarify the role of CDC25B and CDC25C in the development of cancer, and explain the mechanism of tumor cell resistance to radiotherapy and chemotherapy. If the splicing changes of the mRNAs of CDC25B and CDC25C will affect the function of the cells’ checkpoint, the regulation to CDC25B and CDC25C is likely to improve the efficacy of cisplatin. And regulatory pathways and genes will be the new therapeutic targets. Providing important information and evidence for individual treatment of cancer.Objective:To explore the effects of the alternative splicing changes of the mRNA of CDC25B and CDC25C on the growth, proliferation, apoptosis and cisplatin sensitivity of cervical cancer SiHa and C33A cell lines.Methods:The MTT colorimetric method was used to determine the cisplatin semi lethal concentration of the cervical cancer SiHa and C33A cells; RT-PCR was used to detect the effect of the same time, different concentrations of cisplatin intervention and different time, half lethal dose of cisplatin intervention on the alternaive splicing of the mRNA of CDC25B and CDC25C of different groups of cervical cancer cells; By liposome method, CDC25B1, CDC25B3, CDC25C1 and CDC25C5 genes were transfected into cervical cancer SiHa and C33A cells, and thenRT-PCR was used to detect the expression levels of the mRNA of CDC25B1 CDC25B3, CDC25C1 and CDC25C5 in the transfected cells, in addition Western Blot was used to detect the expression levels of the proteins of CDC25B1 CDC25B3, CDC25C1 and CDC25C5 in the transfected cells; The MTT colorimetric method was again used to detect the cisplatin semi lethal concentration of the cervical cancer SiHa and C33A cells with CDC25B1, CDC25B3, CDC25C1 and CDC25C5 over expression. Compared with the previous conclusion and come to the final conclusion.Results:The semi lethal dose of SiHa cells to cisplatin was 40 μmol/L, and the semi lethal dose of C33A cells to cisplatin was 22.5 μmol/L; CDC25B1, CDC25B3, CDC25C1 and CDC25C5, the four most obvious difference spliceosomes, were transfected and up-regulated in cervical cancer cells. And the expression levels of the corresponding mRNAs and proteins, detected by RT-PCR and Western Blot respectively, were up-regulated; The MTT colorimetric method was again used to detect the cisplatin semi lethal concentration of the cervical cancer SiHa and C33A cells with CDC25B1, CDC25B3, CDC25C1 and CDC25C5 over expression. The conclusion compared with the previous conclusion was that, the cisplatin semi lethal concentration was increased significantly in the groups of CDC25B1 and CDC25C1 over expression, and the difference is statistically significant (P< 0.01); the cisplatin semi lethal concentration was decreased significantly in the groups of CDC25B3 and CDC25C5 over expression, and the difference is statistically significant (P< 0.01).Conclusion:CDC25B1, the alternative splicing of the mRNA of CDC25B, can raise the resistance of the cervical cancer SiHa and C33A cell lines to cisplatin CDC25B3, the alternative splicing of the mRNA of CDC25B, can weaken the resistance of the cervical cancer SiHa and C33A cell lines to cisplatin. CDC25C1 the alternative splicing of the mRNA of CDC25C, can raise the resistance of the cervical cancer SiHa and C33A cell lines to cisplatin; CDC25C5, the alternative splicing of the mRNA of CDC25C, can weaken the resistance of the cervical cancer SiHa and C33A cell lines to cisplatin.