本文選題：瞬時(shí)受體電位V6（TRPV6） + 前列腺癌�。� 參考：《南昌大學(xué)》2014年碩士論文

【摘要】：目的： 通過小干擾RNA（siRNA）技術(shù)沉默前列腺癌細(xì)胞中TRPV6基因，初步探討TRPV6對(duì)前列腺癌細(xì)胞增殖和侵襲力的影響及其在前列腺癌雄激素非依賴轉(zhuǎn)化過程中神經(jīng)內(nèi)分泌分化的作用。 方法： （1）體外培養(yǎng)雄激素依賴性前列腺癌細(xì)胞（LNCaP）及雄激素非依賴性前列腺癌細(xì)胞（PC-3），檢測(cè)兩種細(xì)胞中TRPV6的表達(dá)情況。通過siRNA沉默TRPV6基因，檢測(cè)前列腺癌細(xì)胞中TRPV6基因的表達(dá)，，并篩選、鑒定轉(zhuǎn)染穩(wěn)定的LNCaP細(xì)胞。 （2）選取轉(zhuǎn)染成功的LNCaP細(xì)胞并設(shè)定實(shí)驗(yàn)組（轉(zhuǎn)染siRNA-TRPV6）、陰性對(duì)照組（轉(zhuǎn)染siRNA-NC）和空白對(duì)照組（只加轉(zhuǎn)染劑）三組進(jìn)行對(duì)照實(shí)驗(yàn)，通過MTT檢測(cè)細(xì)胞增殖的影響及Transwell檢測(cè)細(xì)胞侵襲力的變化情況，同時(shí)通過熒光定量PCR及western-Blot檢測(cè)神經(jīng)元特異性烯醇化酶（NSE）的表達(dá)情況。 （3）利用SPSS19.0軟件系統(tǒng)建立數(shù)據(jù)庫(kù)和進(jìn)行統(tǒng)計(jì)學(xué)分析，對(duì)比研究TRPV6對(duì)前列腺癌LNCaP細(xì)胞生物學(xué)特征變化的作用。 結(jié)果： 1、熒光定量PCR結(jié)果顯示：前列腺癌雄激素依賴性細(xì)胞株與前列腺癌雄激素非依賴性細(xì)胞株中均存在TRPV6基因的表達(dá)，兩者之間表達(dá)無明顯統(tǒng)計(jì)學(xué)差異（P＞0.05）。 2、轉(zhuǎn)染TRPV6-siRNA后實(shí)驗(yàn)結(jié)果顯示：與陰性對(duì)照組及空白組相比，實(shí)驗(yàn)組的TRPV6呈現(xiàn)低表達(dá)，且有統(tǒng)計(jì)學(xué)意義（P＜0.05）。通過MTT、Transwell等分子生物學(xué)方法檢測(cè)提示：與陰性對(duì)照組及空白組對(duì)比，實(shí)驗(yàn)組細(xì)胞增殖及侵襲力較其他二組均有抑制（P＜0.05），而陰性對(duì)照組與空白組之間則無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 3、熒光定量PCR及western-Blot檢測(cè)實(shí)驗(yàn)組中的神經(jīng)元特異性烯醇化酶（NSE）低于陰性對(duì)照組與空白組（P＜0.05），而陰性對(duì)照組與空白組的表達(dá)無統(tǒng)計(jì)學(xué)差異（P＞0.05），提示TRPV6與前列腺癌雄激素非依賴轉(zhuǎn)化中的神經(jīng)內(nèi)分泌分化存在一定程度正相關(guān)。 結(jié)論： 體外實(shí)驗(yàn)結(jié)果得出沉默TRPV6基因能對(duì)雄激素依賴性前列腺癌LNCaP細(xì)胞的增殖及侵襲力具有抑制作用，并可通過前列腺癌細(xì)胞神經(jīng)內(nèi)分泌分化來影響前列腺癌細(xì)向雄激素非依賴轉(zhuǎn)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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.25

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本文編號(hào)：2019389