前列腺癌雄激素非依赖转化中TRPV6对神经内分泌分化的影响

发布时间：2018-06-14 23:15

本文选题：瞬时受体电位V6（TRPV6） + 前列腺癌　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：通过小干扰RNA（siRNA）技术沉默前列腺癌细胞中TRPV6基因，初步探讨TRPV6对前列腺癌细胞增殖和侵袭力的影响及其在前列腺癌雄激素非依赖转化过程中神经内分泌分化的作用。方法：（1）体外培养雄激素依赖性前列腺癌细胞（LNCaP）及雄激素非依赖性前列腺癌细胞（PC-3），检测两种细胞中TRPV6的表达情况。通过siRNA沉默TRPV6基因，检测前列腺癌细胞中TRPV6基因的表达，，并筛选、鉴定转染稳定的LNCaP细胞。（2）选取转染成功的LNCaP细胞并设定实验组（转染siRNA-TRPV6）、阴性对照组（转染siRNA-NC）和空白对照组（只加转染剂）三组进行对照实验，通过MTT检测细胞增殖的影响及Transwell检测细胞侵袭力的变化情况，同时通过荧光定量PCR及western-Blot检测神经元特异性烯醇化酶（NSE）的表达情况。（3）利用SPSS19.0软件系统建立数据库和进行统计学分析，对比研究TRPV6对前列腺癌LNCaP细胞生物学特征变化的作用。结果： 1、荧光定量PCR结果显示：前列腺癌雄激素依赖性细胞株与前列腺癌雄激素非依赖性细胞株中均存在TRPV6基因的表达，两者之间表达无明显统计学差异（P＞0.05）。 2、转染TRPV6-siRNA后实验结果显示：与阴性对照组及空白组相比，实验组的TRPV6呈现低表达，且有统计学意义（P＜0.05）。通过MTT、Transwell等分子生物学方法检测提示：与阴性对照组及空白组对比，实验组细胞增殖及侵袭力较其他二组均有抑制（P＜0.05），而阴性对照组与空白组之间则无统计学差异（P＞0.05）。 3、荧光定量PCR及western-Blot检测实验组中的神经元特异性烯醇化酶（NSE）低于阴性对照组与空白组（P＜0.05），而阴性对照组与空白组的表达无统计学差异（P＞0.05），提示TRPV6与前列腺癌雄激素非依赖转化中的神经内分泌分化存在一定程度正相关。结论：体外实验结果得出沉默TRPV6基因能对雄激素依赖性前列腺癌LNCaP细胞的增殖及侵袭力具有抑制作用，并可通过前列腺癌细胞神经内分泌分化来影响前列腺癌细向雄激素非依赖转化。
[Abstract]:Objective: to silence TRPV6 gene in prostate cancer cells by small interference RNA-siRNAs. To explore the effect of TRPV6 on the proliferation and invasion of prostate cancer cells and the role of neuroendocrine differentiation in the process of androgen independent transformation of prostate cancer. Methods: 1) androgen dependent prostate cancer cell line LNCaP) and androgen independent prostate cancer cell line (PC-3) were cultured in vitro to detect the expression of TRPV6. The expression of TRPV6 gene in prostate cancer cells was detected by silencing TRPV6 gene with siRNA. The transfected LNCaP cells were selected and the three groups (transfected siRNA-TRPV6, negative control group (transfected siRNA-NCX) and blank control group (only with transfection agent) were set up to carry out the control experiment. MTT assay was used to detect the effect of cell proliferation and Transwell was used to detect the changes of cell invasiveness. Meanwhile, the expression of neuron-specific enolase (NSE) was detected by fluorescent quantitative PCR and western-Blot. To study the effect of TRPV6 on the biological characteristics of prostate cancer LNCaP cells. Results: 1. The results of fluorescence quantitative PCR showed that TRPV6 gene was expressed in androgen-dependent and androgen-independent cell lines of prostate cancer. There was no significant difference in the expression of TRPV6 between the two groups (P > 0.05). After transfection of TRPV6-siRNA, the results showed that the expression of TRPV6 in the experimental group was lower than that in the negative control group and the blank group (P < 0.05). The results of molecular biological methods such as MTT / Transwell showed that: compared with the negative control group and the blank group, Compared with the other two groups, the proliferation and invasiveness of the experimental group were inhibited (P < 0.05), but there was no significant difference between the negative control group and the blank group (P > 0.05). The neuronal specificity in the experimental group was detected by fluorescence quantitative PCR and Western-Blot. The expression of enolase NSEwas lower than that of negative control group and blank group (P < 0.05), but there was no significant difference between negative control group and blank group (P > 0.05), suggesting that TRPV6 was positively correlated with neuroendocrine differentiation in androgen independent transformation of prostate cancer. Conclusion: silencing TRPV6 gene can inhibit the proliferation and invasion of androgen dependent prostate cancer LNCaP cells in vitro. The differentiation of prostate cancer cells into androgen-independent transformation can be effected by neuroendocrine differentiation of prostate cancer cells.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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