| Background:Ovarian cancer is one of the three common female reproductive malignanttumors with the highest mortality among them. The mortality caused by itranked the fifth in the ten leading types of malignant tumors in female with theannual mortality around of9.5in10thousand[1]. Therefore, ovarian cancerseriously threatens women’s health and life. Operation is the preferred treatmentof ovarian cancer, postoperative supplemented by therapy or radiation chemicaldrug treatment for conventional therapy. However, because of side effects of thechemotherapy medicines or their dissolution, the expensive fee, and primary oracquired drug resistance, the chemical treatment is often difficult to achieve theideal effect. The drug resistance of tumor cells to chemotherapy drugs isseriously affecting the effects of ovarian cancer chemotherapy and survivalquality, results in the5-year survival rate of patients drops greatly, which is oneof the main cause of failure treatment[2]. Therefore, mining new anti-cancer drug,low toxicity, high efficiency or chemo-therapy sensitizing drugs is the key toimprove the prognosis of ovarian cancer.Resveratrol (nods Res) is compound of containg stilbene type structure offlavonoid polyphenol which can be found in grapes, liliaceous, leguminosae, polygonaceae and other more than70kinds of plants. It posseses anti inflamm-atorys, antibacterial, antioxidant, antivirus, resistance to atherosclerosis, immuneregulation, and a variety of biological functions such as nerve protection. Andone of the most important pharmacological effects is Bai Li reed alcohol canpromote apoptosis and inhibit the growth of tumor cell proliferation throughvarious pathways. It has been reported that Bai Li reed alcohol has a therapeuticeffects on liver cancer, ovarian cancer, leukemia, laryngeal squamous carcinoma,breast cancer, prostate cancer, thyroid cancer, and skin squamous carcinoma[3].Res is regarded as a potential tumor inhibitor, there is more research onleukemia, breast cancer, liver cancer, lung cancer, colorectal cancer, cervicalcancer and others, while research on ovarian cancer is rare at home and abroad.Therefore, studying the effect of resveratrol on cell proliferation and apoptosisof ovarian cancer, and exploring the mechanism of apoptosis, has importantclinical value in order to provide a new breakthrough in the prevention andtreatment of ovarian cancer.Objective:To study the resveratrol induced apoptosis of human ovarian cancer cellsSKOV-3, to explore the mechanism of apoptosis. Resveratrol is expected tobecome new ovarian cancer treatments and we expect to find its targets for EOCand provide its theory in treating EOC in clinic.Methods: 1. Cell culture: the recovery SKOV-3ovarian cancer cells, the vaccinationin RPMI1640culture medium containing10%calf serum, at37℃cellscontaining5%CO2incubator in the training.2.5g/L trypsin and0.2g/L EDTAdigestion, extend;2. Cell proliferation inhibition (determined by MTT reduction method):composite nutrient solution with different concentrations of resveratrol(including400umol/L,200umol/L,100umol/L,50umol/L,25umol/L), to acton human ovarian cancer SKOV-3cells, by determined by MTT method todetect resveratrol effect on ovarian cancer SKOV-3cell proliferation;3. Transmission electron microscopy (tem): by observing the instrumentcontrol group, in the role of different concentrations of resveratrol group afterSKOV-3ovarian cancer cells, cell ultrastructure change;4. Flow cytometry analysis: the composite nutrient solution culture withdifferent concentrations of resveratrol48hours SKOV-3people ovarian cancercells, cells in the quantitative analysis of each group of changes of cell cycle andcell apoptosis;5. RT-PCR method: SKOV-3people ovarian cancer cells were dividedinto different concentrations of resveratrol48hours, using RT-PCR methoddetected the relative mRNA levels and expression of the bcl-2and bax.6. Westernblot method: SKOV-3people ovarian cancer cells were dividedinto different concentrations of resveratrol48hours, using Westernblot methoddetected the relative protein levels and expression of the bcl-2and bax. Results:1. Different concentrations of resveratrol have effect of inhibition onSKOV-3ovarian cancer cells with concentration and time dependence.2. The changes of typical apoptotic cells can be observed under thetransmission electron microscopy (tem) in different concentration of resveratrolSKOV-3groups.3. The results from flow cytometer shows that cell apoptosis rate increasedand the cell cycle changed obviously with the increased concentration ofresveratrol in48hours:the percentage of G0/G1phase cell decreased obviously,the percentage of S phase cell inceased, which indicated that resveratrol inducedcell apoptosis by inhibiting cell DNA synthesis.4. RT-PCR detecting the Bcl-2and Bax mRNA expression: the positiveresults of RT-PCR to detect SKOV-3Bcl-2cells and Bax mRNA expressionwere dectected with different concentrations of resveratrol (25,50,100,200，400umol/L) after48h, and the Bcl-2mRNA stripe density was decreased withthe increase of concentration, Bax mRNA stripe density was increased with theincreased of the concentration. The results show that resveratrol induced cellapoptosis probably by cutting the Bcl-2expression and increasing the expressionof Bax.5. Westernblot detecting the Bcl-2and Bax protein expression: undercondition of25,50,100,200，400umol/L resveratrol action SKOV-3cell48h,the Bcl-2protein with tapering became thiner but Bax protein gradually became thicker with the increased of the concentration of resveratrol. The results showthat resveratrol induced cell apoptosis probably by cutting the Bcl-2expressionand increasing the expression of Bax.Conclusion:1. Resveratrol can inhibit SKOV-3ovarian cancer cell proliferation andinduce its apoptosis;2. Resveratrol can block SKOV-3ovarian cancer cell proliferation in Sphase;3. The mechanism of resveratrol inducing apoptosis of ovarian cancerSKOV-3cells might be related to the inhibition of antiapoptosis factorexpression of Bcl-2and enhancement of the expression apoptosis factor of Bax. |
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