| Background:Cervical cancer is a common disease of women, which the incidence rate after breast cancer ranks second in gynecological malignancies, and serious threats women’s lives and health. In recent years, the incidence of cervical cancer is gradually increased, and more and more young.The pathogenesis of cervical cancer is unclear, previous treatment for cervical cancer mainly by operation, chemotherapy, radiotherapy and biological related treatment with HPV vaccines, but all of them have its own limitations. Therefore, to develop safe and effective anticancer drugs has become the main direction of researchers.Aspirin is one of a kind of Non-steroidal anti-inflammatory drug (NSAID), it is also called as acetylsalicylic acid (ASA). Aspirin is a inhibitor of cyclooxygenase and it is widely used in antipyretic, analgesic and anti-inflammatory for a long time. Recent studies have shown that aspirin can be used as a drug which can treat and prevent cancer, and may enhance the anti-tumor effect in combination with chemotherapy drugs such as cisplatin, which for the treatment and rehabilitation process of cancer patients provides a new selection. However, the treatment also exist a great risk because the antitumor efficacy of aspirin is not high enough.Triptolide (TPL) is the mixture that the major composition extracted from the Chinese herb Tripteryglum wilfordii Hook.f, In the study, the main traditional role of Triptolide is anti-inflammatory, immunosuppressive, and anti-fertility etc. Now, it has been proved that triptolide can inhibit the growth of tumor, and including higher antitumor effect. However, careful consideration needs when in application because of some side effects of itself.About the controversary that how to reduce the potential side effects of triptolide and turely applicatted in clinical application, it is an urgent problem to be solved.The combined treatment of traditional Chinese medicine prescription, as in recent years, for the treatment of cancer has become a popular trend inthe use of combined traditional Chinese medicine and Western medicine,because of non-steroidal anti-inflammatory drug aspirin and traditional Chinese medicine derivative of triptolide has similar effects in the treatment of cervical cancer, because of this, this study based on the past research and the cervical cancer cells as the research object, analyze and study the effect and efficiency of Tripterygium Wilfordii and non-steroidal anti-inflammatory drug aspirin in invasion and apoptosis of inmalignant features in cervical cancer, and research its relevant mechanisms, this will provide a new strategy for the treatment and prevention of cervical cancer.Objective:Aim of this study was to analyze the anti-tumor affect and efficiency of the non-steroidal anti-inflammatory drug aspirin of aspirin and the traditional Chinese medicine derivative of triptolide in cervical cancer cells, and to provide a strategy to therapy and prevetion of cervical cancer.Methods:In the study, in the first place, viability of cervical cancer cell lines was assessed by the MTT method at various concentrations of aspirin (5-12.5μM) and triptolide (50-125 nM)in the condition of the drug independent and combination.In the second place, apoptosis fuction of cervical cancer cell lines was assessed by the methods of Flow Cytometry and Mitochondrial Membrane Detection at various concentrations of aspirin (5-12.5μM) and triptolide (50-125 nM) in the condition of the drug independent and combination, meanwhile, apoptosis of cervical cancer cell lines was assessed by the methods of the Clone Formation Assay at only concentrations of aspirin (10μM) and triptolide (100 nM) in the condition of the drug independent and combination. In the last place, the technology of quantitation Real time-PCR and Western Blotting were used to analyze the expression of the cell cycle related factor of Cyclin E and p21 as the cell cycle apotosis factor of Bcl-2 and Bax after that only concentrations of aspirin(10μM) and triptolide (100nM) was used in the condition of the drug independent and combination.Result:The result of this study indicates that viability in the combination group by the method of MTT was significantly decreased as compared with either drug used alone and shown apperantly the effect of synergistic anti-tumor on cervical cancer cells(P< 0.01), meanwhile, the fuction of suppressing the proliferation with a dose-dependent manner in cervical cancer cell line. Apoptosis function in the combination group by the method of Flow Cytometry membrane was significantly increased as compared with either drug used alone(P< 0.05), meanwhile, the fluorescence intensity by the method of Mitochondrial Membrane Detection was significantly decreased as compared with either drug used alone(P< 0.05). In addition, the number of clone formation by the method of Clone Formation Assay was significantly decreased as compared with either drug used alone(P< 0.05). Furthermore, the results by the method of qRT-PCR and Western bolt shown that the expression of the Cyclin E and Bcl-2 in the combination group was significantly decreased as compared with either drug used alone(P< 0.05), nevertheless, the expression of the p21 and Bax in the combination group was significantly increased as compared with either drug used alone(P<0.05).Conclusion:In conclusion, we can summarize from these results, first of all, asprin and triptolide not only have anti-proliferation effect t on the growth of cervical cancer cells used single, but have the synergistic antiproliferative effects when used together. in the second, apoptosis percentage of cervical cancer cell lines increased significantly in the condition of the drug combination than independent. At the same time, aspirin combined with triptolide can induce the Mitochondrial membrane potential significantly decreased (fluorescence intensity);in the third, the combination of aspirin and triptolide can inhibit the ability of cervical cancer cell colony forming;in the last, the combination of aspirin and triptolide can affect the expression levels of the cell cycle and apoptosis related factors changing significantly. |
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