【摘要】：目的： 通過觀察姜黃素(curcumin)對人卵巢癌A2780/Taxol細胞作用，研究細胞耐藥的原因及姜黃素逆轉(zhuǎn)耐藥的機制，為姜黃素逆轉(zhuǎn)卵巢癌耐藥應(yīng)用于臨床提供實驗依據(jù)。 方法： 1.卵巢癌耐藥細胞A2780/Taxol在體外培養(yǎng)，MTT比色法檢測不同濃度的姜黃素對細胞增殖的影響。 2. Western-blot方法檢測姜黃素作用人卵巢癌A2780/Taxol細胞后，細胞內(nèi)P-gP，PKC-α的表達情況。 3.姜黃素、紫杉醇共同作用于卵巢癌耐藥細胞A2780/Taxol，經(jīng)PI（碘化丙啶）染色后，上流式細胞儀測定細胞凋亡情況。 結(jié)果： 1.紫杉醇與不同濃度姜黃素聯(lián)合作用于卵巢癌耐藥細胞A2780/Taxol，較對照組能明顯抑制細胞生長（P0.01）；姜黃素濃度未達20μM時，伴隨姜黃素濃度的遞增，人卵巢癌A2780/Taxol細胞生長抑制呈上升趨勢，差異具有統(tǒng)計學意義（P0.01）；姜黃素濃度達20μM后，姜黃素濃度對卵巢癌A2780/Taxol細胞的生長抑制率無明顯影響（P0.05）。 2. western blot結(jié)果表明姜黃素干預(yù)的人卵巢癌A2780/Taxol細胞，Mdr-1/P-gP、PKC-α蛋白表達比對照組明顯降低，隨著姜黃素濃度的增加Mdr-1/P-gP、PKC-α蛋白的表達有明顯下降趨勢，當姜黃素濃度為增加到20μM時隨著姜黃素濃度的增加Mdr-1/P-gP、PKC-α蛋白的表達不再下降。 3.不同組細胞PI染色后，上流式細胞儀檢測，結(jié)果發(fā)現(xiàn)姜黃素組細胞的凋亡率明顯比空白對照組增加（P0.05），姜黃素濃度未達20μM時，伴隨姜黃素濃度的遞增人卵巢癌A2780/Taxol細胞的凋亡率有上升趨勢,差異具有統(tǒng)計學意義（P0.01），姜黃素濃度達20μM后，姜黃素濃度對卵巢癌A2780/Taxol細胞的生長抑制率無明顯關(guān)系，差異無統(tǒng)計學意義（P0.05）。姜黃素20μM聯(lián)合紫杉醇組凋亡率比單獨使用紫杉醇組明顯升高，差異具有統(tǒng)計學意義（P0.01）。 結(jié)論: 1、姜黃素能抑制人卵巢癌A2780/Taxol細胞的生長，具有抗腫瘤作用，姜黃素與紫杉醇聯(lián)合作用較單獨使用紫杉醇細胞抑制率明顯增加，姜黃素能增加紫杉醇對人卵巢癌A2780/Taxol細胞的細胞毒性，降低細胞的耐藥性。 2、姜黃素能通過抑制人卵巢癌A2780/Taxol中Mdr-1基因表達，降低P-gP蛋白表達，阻礙P-gP蛋白生物泵功能，減少卵巢癌細胞對紫杉醇的泵除，增加細胞對紫杉醇敏感性。 3、姜黃素可以降低人卵巢A2780/Taxol細胞中PKC-α蛋白的表達，降低P-gp的磷酸化速度，抑制P-gP蛋白生物學功能，逆轉(zhuǎn)P-gP介導的卵巢癌耐藥。
[Abstract]:Objective: to observe the effect of curcumin (curcumin) on human ovarian cancer A2780/Taxol cells, and to study the cause of cell resistance and the mechanism of curcumin reversing drug resistance in order to provide experimental evidence for curcumin reversal of ovarian cancer resistance in clinic. Methods: 1. Ovarian cancer cell line A2780/Taxol was cultured in vitro. The effect of curcumin at different concentrations on cell proliferation was detected by MTT colorimetry. 2. The expression of P-gPtPKC- 偽 in human ovarian cancer A2780/Taxol cells was detected by Western-blot. 3. Curcumin and paclitaxel acted on ovarian cancer cell line A2780 / Taxol.After PI staining, apoptosis was detected by upflow cytometry. Results: 1. Paclitaxel combined with curcumin at different concentrations could inhibit the growth of ovarian cancer cell line A2780 / Taxol.Compared with the control group (P0.01), paclitaxel could inhibit the growth of ovarian cancer cells significantly (P0.01). When curcumin concentration was not up to 20 渭 M, the growth inhibition of human ovarian cancer A2780/Taxol cells increased with the increase of curcumin concentration, and the difference was statistically significant (P0.01). When curcumin concentration reached 20 渭 M, curcumin concentration had no significant effect on growth inhibition rate of ovarian cancer A2780/Taxol cells (P0.05). 2. The results of western blot showed that the expression of Mdr-1/P-gP,PKC- 偽 protein in A2780/Taxol cells of human ovarian cancer treated with curcumin was significantly lower than that in control group, and Mdr-1/P-gP, was increased with the increase of curcumin concentration. The expression of PKC- 偽 protein decreased obviously. When the concentration of curcumin increased to 20 渭 M, the expression of Mdr-1/P-gP,PKC- 偽 protein did not decrease with the increase of curcumin concentration. 3. After PI staining, the apoptosis rate of curcumin group was significantly higher than that of control group (P0.05), and when curcumin concentration was less than 20 渭 M, the apoptosis rate of curcumin group was significantly higher than that of control group (P0.05). The apoptosis rate of human ovarian cancer A2780/Taxol cells increased with increasing curcumin concentration, and the difference was statistically significant (P0.01). When curcumin concentration reached 20 渭 M, the apoptosis rate of human ovarian cancer A2780/Taxol cells increased significantly. The concentration of curcumin had no significant relationship with the growth inhibition rate of ovarian cancer A2780/Taxol cells (P0.05). The apoptosis rate of 20 渭 M curcumin combined with paclitaxel group was significantly higher than that of paclitaxel alone group (P0.01). Conclusion: 1. Curcumin can inhibit the growth of human ovarian cancer A2780/Taxol cells and has anti-tumor effect. The inhibitory rate of curcumin combined with paclitaxel is significantly higher than that of paclitaxel alone. Curcumin can increase the cytotoxicity of paclitaxel to human ovarian cancer A2780/Taxol cells and reduce cell resistance. 2. Curcumin can inhibit the expression of Mdr-1 gene, decrease the expression of P-gP protein, block the biological pump function of P-gP protein, reduce the pump removal of paclitaxel and increase the sensitivity of ovarian cancer cell to paclitaxel by inhibiting the expression of Mdr-1 gene in human ovarian cancer A2780/Taxol. 3. Curcumin could decrease the expression of PKC- 偽 protein, decrease the phosphorylation rate of P-gp, inhibit the biological function of P-gP protein, and reverse the drug resistance of ovarian cancer mediated by P-gP.
【學位授予單位】：蚌埠醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.31

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