姜黄素逆转卵巢癌耐药的试验研究
发布时间：2019-01-01 14:02
【摘要】：目的： 通过观察姜黄素(curcumin)对人卵巢癌A2780/Taxol细胞作用，研究细胞耐药的原因及姜黄素逆转耐药的机制，为姜黄素逆转卵巢癌耐药应用于临床提供实验依据。 方法： 1.卵巢癌耐药细胞A2780/Taxol在体外培养，MTT比色法检测不同浓度的姜黄素对细胞增殖的影响。 2. Western-blot方法检测姜黄素作用人卵巢癌A2780/Taxol细胞后，细胞内P-gP，PKC-α的表达情况。 3.姜黄素、紫杉醇共同作用于卵巢癌耐药细胞A2780/Taxol，经PI（碘化丙啶）染色后，上流式细胞仪测定细胞凋亡情况。 结果： 1.紫杉醇与不同浓度姜黄素联合作用于卵巢癌耐药细胞A2780/Taxol，较对照组能明显抑制细胞生长（P0.01）；姜黄素浓度未达20μM时，伴随姜黄素浓度的递增，人卵巢癌A2780/Taxol细胞生长抑制呈上升趋势，差异具有统计学意义（P0.01）；姜黄素浓度达20μM后，姜黄素浓度对卵巢癌A2780/Taxol细胞的生长抑制率无明显影响（P0.05）。 2. western blot结果表明姜黄素干预的人卵巢癌A2780/Taxol细胞，Mdr-1/P-gP、PKC-α蛋白表达比对照组明显降低，随着姜黄素浓度的增加Mdr-1/P-gP、PKC-α蛋白的表达有明显下降趋势，当姜黄素浓度为增加到20μM时随着姜黄素浓度的增加Mdr-1/P-gP、PKC-α蛋白的表达不再下降。 3.不同组细胞PI染色后，上流式细胞仪检测，结果发现姜黄素组细胞的凋亡率明显比空白对照组增加（P0.05），姜黄素浓度未达20μM时，伴随姜黄素浓度的递增人卵巢癌A2780/Taxol细胞的凋亡率有上升趋势,差异具有统计学意义（P0.01），姜黄素浓度达20μM后，姜黄素浓度对卵巢癌A2780/Taxol细胞的生长抑制率无明显关系，差异无统计学意义（P0.05）。姜黄素20μM联合紫杉醇组凋亡率比单独使用紫杉醇组明显升高，差异具有统计学意义（P0.01）。 结论: 1、姜黄素能抑制人卵巢癌A2780/Taxol细胞的生长，具有抗肿瘤作用，姜黄素与紫杉醇联合作用较单独使用紫杉醇细胞抑制率明显增加，姜黄素能增加紫杉醇对人卵巢癌A2780/Taxol细胞的细胞毒性，降低细胞的耐药性。 2、姜黄素能通过抑制人卵巢癌A2780/Taxol中Mdr-1基因表达，降低P-gP蛋白表达，阻碍P-gP蛋白生物泵功能，减少卵巢癌细胞对紫杉醇的泵除，增加细胞对紫杉醇敏感性。 3、姜黄素可以降低人卵巢A2780/Taxol细胞中PKC-α蛋白的表达，降低P-gp的磷酸化速度，抑制P-gP蛋白生物学功能，逆转P-gP介导的卵巢癌耐药。
[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- 伪 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- 伪 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- 伪 protein decreased obviously. When the concentration of curcumin increased to 20 渭 M, the expression of Mdr-1/P-gP,PKC- 伪 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- 伪 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.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
本文编号：2397670
[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- 伪 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- 伪 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- 伪 protein decreased obviously. When the concentration of curcumin increased to 20 渭 M, the expression of Mdr-1/P-gP,PKC- 伪 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- 伪 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.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
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