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Objective: research manumycin (Manumycin) combined with cisplatin (DDP) 3AO vitro inhibition of human ovarian cancer, and inhibition of nude mice, analysis manumycin plus cisplatin inhibited 3AO proliferation and induced withered. The perish possible mechanism to explore the feasibility of of manumycin used in combination with cisplatin, ovarian cancer. Methods: 1 in order to the manumycin effect of different concentrations in vitro culture of human ovarian cancer 3AO cells tetramethylazodicarboxamide tetrazolium (tetrzolium-based colorimetric assay, MTT) colorimetric test detects cell proliferation activity, and the cells were observed The morphological changes. MTT colorimetric assay manumycin combined with cisplatin on 3AO proliferation inhibition, the use of the Guinness Book of formula [1] analysis of the interaction index (Iteraction Index), evaluation of joint, such as q gt; 0.85 indicates relative plus the role, q lt; 0.85 antagonism. 3 establish 3AO cells transplanted into nude mice model, two weeks after the tumor grew to the vaccination, were randomly divided into four groups, the manumycin group (n = 5): control group, cisplatin group, the combination group. Nude mice such as the diet, activity, and the reaction course of the experiment. Growth record in each group transplanted into nude mice, tumor growth curve to calculate the rate of tumor growth inhibition. Immunohistochemistry assay in tumor tissues of Survivin, NF-κB, VEGF, Caspase-3 protein expression. Flow cytometry (flow cytometry, FCM) to detect changes in the transplanted tumor tissue cell cycle and apoptosis rate. Results: 1 MTT colorimetric 3AO proliferation manumycin (5,10,20,40,60 μmol / L) inhibited. Different concentrations of manumycin-treated cells 24 ~ 72h, compared with the control group, the OD value of each treatment group were decreased, and the difference was statistically significant (p lt; 0.05). Readily neomycin concentration increases, the extension of the duration of action OD values ??decreased manumycin 3AO cell proliferation inhibition is dose-dependent effect was obvious - the time. Cell morphology was observed under light microscope: fusiform cells without drug action, shape and full, while the drug group cell shrinkage, cracking, irregular-shaped, exfoliated cells increased inhibition 3AO cell growth. 2 MTT colorimetric analysis manumycin plus cisplatin 3AO cell proliferation inhibition results showed: 10, 20, 40, μmol / L manumycin enhanced the inhibition of cisplatin 3AO cell proliferation, but also readily neomycin increase concentration, enhance its role in combination with cisplatin. Guinness formula to analyze the interaction index showed that different concentrations of cisplatin combined with 10, 20, 40, μmol / L manumycin q value gt; 0.85. 3 successfully established ovarian carcinoma xenografts in nude mice inoculated success rate of 100% . Nude mice transplanted tumor volume after the end of treatment, the treatment groups than the control group, the combined treatment group is less than the monotherapy group, the differences were statistically significant (p lt; 0.05). The control group the manumycin group and cisplatin group, the combination group, tumor inhibition rate was 0.00%, 29.85%, 37.36%, and 64.60%, respectively. 3AO cell xenograft tumor tissue immunohistochemistry assay Survivin, VEGF, NF-Κb, Caspase-3 protein expression: immunohistochemistry score for each indicator, each treatment group and the control group compared to the combination group and compared to the monotherapy group, the differences were statistically significant (p lt; 0.05). Flow cytometry results showed that: the control group the manumycin group, cisplatin group, the combination group, cell apoptosis rate gradually increased to 5.24 ± 0.94%, respectively, 12.16 ± 1.08%, 17.41 ± 2.26%, 22.93 ± 3.22%, the cells in G0/G1 phase of the cell cycle is gradually reduced, the G2 / M phase cells gradually increased, S phase cells did not change significantly. Conclusion: the 1 Manumycin 3AO cell proliferation inhibition was time - a dose-dependent relationship, the manumycin cisplatin and cisplatin 3AO cell proliferation inhibition. 3AO cell xenografts in nude mice 2 manumycin can inhibit the growth of manumycin cisplatin enhanced antitumor effect. 3 manumycin allows 3AO cell, the cell cycle arrest at the G2 / M phase and induce apoptosis 3AO cell manumycin cisplatin-induced apoptosis enhanced role. 4 manumycin increased Caspase-3 protein expression by inhibiting NF-κB, VEGF, Survivin protein expression, enhanced 3AO cell sensitivity to cisplatin.
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