雷公藤红素在前列腺癌细胞中增敏多西紫杉醇的研究

发布时间：2018-06-08 02:22

  本文选题：雷公藤红素 + 多西紫杉醇　； 参考：《哈尔滨工业大学》2016年硕士论文

【摘要】：前列腺癌(Prostate Cancer,PCa)是在男性中发病率和死亡率都非常高的癌症之一。虽然随着检测手段的更新以及治疗方式的多样化,但是大部分病人一旦发展出转移瘤,进而转变为CRPC(Castration-resistant PCa),最终产生药物抗性时,其生存期将大大降低。CRPC被认为与AR(Androgen Receptor)通路的再激活关系密切,AR通路的再激活是导致肿瘤存活的关键。AR剪切变异体AR-V7不需要雄激素激活转运入核,能够激活AR通路调控下游基因。因此,AR-V7已成为治疗CRPC前列腺癌的肿瘤治疗的研究热点。多西紫杉醇作为治疗CRPC的一线化疗药物,能够通过抑制微管解聚进而抑制AR核转入并激活细胞凋亡。然而CRPC最终会发展出药物抗性,抗性来源可能是凋亡通路的改变以及AR-V7的表达。雷公藤红素作为一种天然的蛋白酶体抑制剂,能够通过诱导细胞凋亡、抑制AR的表达进而抑制前列腺癌细胞系的生长。因此,本研究利用AR-V7阳性细胞系22Rv1作为模型,并将AR-V7阴性细胞系LNCa P作为对照,分析并解释多西紫杉醇与雷公藤红素对前列腺癌细胞的生长抑制作用差异以及其机理。本研究通过MTT实验确证了22Rv1为多西紫杉醇抗性细胞系、LNCa P则为敏感细胞系,同时与雷公藤红素联合用药后细胞存活率相似,联合用药计算显示雷公藤红素能够恢复22Rv1对多西紫杉醇的敏感性,而LNCa P经过药物联用处理后则表现出拮抗效果。凋亡实验显示,0.5 nmol/L的多西紫杉醇与0.8μmol/L的雷公藤红素单独用药对AR-V7表达差异的两株前列腺癌细胞诱导的凋亡结果无明显差异,药物联用下22Rv1的凋亡率上升至50.62%,LNCa P则仅为32.6%。这说明了雷公藤红素在AR-V7阳性细胞系中能恢复多西紫杉醇诱导凋亡的能力。同时Western Blot结果显示,雷公藤红素能够恢复多西紫杉醇在多西紫杉醇抗性细胞系中的PARP切割,并且在多西紫杉醇处理下22Rv1细胞系较LNCa P细胞PARP切割表达量少,这说明了雷公藤红素能够通过恢复多西紫杉醇抗性细胞系内已改变的凋亡通路产生增敏作用。q RT-PCR结果显示,雷公藤红素能够从m RNA水平上抑制AR、AR-V7的表达,而多西紫杉醇不能够影响其表达。这说明,雷公藤红素能够通过抑制AR、AR-V7表达抑制细胞内的AR通路。综上,本研究初步分析雷公藤红素与多西紫杉醇对前列腺癌细胞生长抑制差异,阐明雷公藤红素抑制AR-V7阳性细胞生长、药物增敏的部分分子机制。
[Abstract]:Prostate Cancer (PCa) is one of the cancers with high morbidity and mortality among men. Although with the renewal of detection methods and the diversification of the methods of treatment, the survival period will be large when most patients develop metastatic tumors and then turn to CRPC (Castration-resistant PCa) and eventually produce drug resistance. .CRPC is believed to be closely related to the reactivation of the AR (Androgen Receptor) pathway, and the reactivation of the AR pathway is the key.AR shear variant AR-V7 that does not require androgen activation to enter the nucleus, and can activate the AR pathway to regulate the downstream genes. Therefore, AR-V7 has become a research heat for the treatment of cancer of CRPC prostate cancer. As a first-line chemotherapeutic agent for the treatment of CRPC, docetaxel can inhibit microtubule disaggregation and inhibit the transfer of AR nuclei to activate cell apoptosis. However, CRPC eventually develops drug resistance, and the source of resistance may be a change in the apoptosis pathway and the expression of AR-V7. In this study, the AR-V7 positive cell line 22Rv1 was used as a model, and the AR-V7 negative cell line LNCa P was used as the control, and the difference in the inhibitory effect of docetaxel and tripterin on the growth of the adenocarcinoma cells of the anterior adenocarcinoma cells was analyzed and explained, as well as the inhibition of the growth of the prostate cancer cell lines by inducing apoptosis. Therefore, the AR positive cell line 22Rv1 was used as a model. Mechanism. In this study, the MTT test confirmed that 22Rv1 was a docetaxel resistant cell line, LNCa P was a sensitive cell line, and the cell survival rate was similar to that of tripterin. The combined use of tripterin could restore the sensitivity of 22Rv1 to docetaxel, while LNCa P was treated with a combination of drugs. The apoptosis test showed that 0.5 nmol/L of docetaxel and 0.8 mol/L of tripterin alone had no significant difference in the apoptosis induced by two prostate cancer cells with different AR-V7 expression, the apoptosis rate of 22Rv1 increased to 50.62%, and LNCa P was only 32.6%., which indicated that tripterin in AR-V7 In the positive cell line, the ability to induce docetaxel to induce apoptosis was restored. Western Blot results showed that tripterine could restore the PARP cutting of docetaxel in the docetaxel resistant cell line, and the PARP cut expression in 22Rv1 cell lines was less than that of LNCa P cells under docetaxel treatment, which indicated that tripterine was reproduced. .q RT-PCR can be produced by restoring the altered apoptotic pathway in the cell line of the docetaxel resistant cell line. The results show that Lei Gongteng erythropoietin can inhibit the expression of AR and AR-V7 from m RNA level, while docetaxel can not affect its expression. This shows that Lei Gongteng erythropoietin can inhibit AR by inhibiting AR and AR-V7 expression. To sum up, this study preliminarily analyzed the difference between tripterine and docetaxel on the growth inhibition of prostate cancer cells, and elucidated the molecular mechanism of tripterine inhibiting the growth of AR-V7 positive cells and sensitizing the drugs.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R737.25
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本文编号：1993973