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

【摘要】：目的： 通過小干擾RNA（siRNA）技術沉默前列腺癌細胞中TRPV6基因，初步探討TRPV6對前列腺癌細胞增殖和侵襲力的影響及其在前列腺癌雄激素非依賴轉化過程中神經內分泌分化的作用。 方法： （1）體外培養(yǎng)雄激素依賴性前列腺癌細胞（LNCaP）及雄激素非依賴性前列腺癌細胞（PC-3），檢測兩種細胞中TRPV6的表達情況。通過siRNA沉默TRPV6基因，檢測前列腺癌細胞中TRPV6基因的表達，，并篩選、鑒定轉染穩(wěn)定的LNCaP細胞。 （2）選取轉染成功的LNCaP細胞并設定實驗組（轉染siRNA-TRPV6）、陰性對照組（轉染siRNA-NC）和空白對照組（只加轉染劑）三組進行對照實驗，通過MTT檢測細胞增殖的影響及Transwell檢測細胞侵襲力的變化情況，同時通過熒光定量PCR及western-Blot檢測神經元特異性烯醇化酶（NSE）的表達情況。 （3）利用SPSS19.0軟件系統(tǒng)建立數(shù)據(jù)庫和進行統(tǒng)計學分析，對比研究TRPV6對前列腺癌LNCaP細胞生物學特征變化的作用。 結果： 1、熒光定量PCR結果顯示：前列腺癌雄激素依賴性細胞株與前列腺癌雄激素非依賴性細胞株中均存在TRPV6基因的表達，兩者之間表達無明顯統(tǒng)計學差異（P＞0.05）。 2、轉染TRPV6-siRNA后實驗結果顯示：與陰性對照組及空白組相比，實驗組的TRPV6呈現(xiàn)低表達，且有統(tǒng)計學意義（P＜0.05）。通過MTT、Transwell等分子生物學方法檢測提示：與陰性對照組及空白組對比，實驗組細胞增殖及侵襲力較其他二組均有抑制（P＜0.05），而陰性對照組與空白組之間則無統(tǒng)計學差異（P＞0.05）。 3、熒光定量PCR及western-Blot檢測實驗組中的神經元特異性烯醇化酶（NSE）低于陰性對照組與空白組（P＜0.05），而陰性對照組與空白組的表達無統(tǒng)計學差異（P＞0.05），提示TRPV6與前列腺癌雄激素非依賴轉化中的神經內分泌分化存在一定程度正相關。 結論： 體外實驗結果得出沉默TRPV6基因能對雄激素依賴性前列腺癌LNCaP細胞的增殖及侵襲力具有抑制作用，并可通過前列腺癌細胞神經內分泌分化來影響前列腺癌細向雄激素非依賴轉化。
[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.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.25

【參考文獻】

相關期刊論文 前4條

1 趙曉智;郭宏騫;劉光香;紀長威;張士偉;劉鐵石;甘衛(wèi)東;李笑弓;;靶向TRPV6的siRNA對前列腺癌LNCaP細胞抑制作用的體外研究[J];中華男科學雜志;2010年05期

2 史娟,李繼碩;TRP離子通道[J];神經解剖學雜志;2004年02期

3 夏術階,唐孝達;前列腺導管系統(tǒng)遠近端的細胞凋亡、雄激素受體亞型與雄雌激素環(huán)境研究[J];中華實驗外科雜志;2003年02期

4 宋毅,吳關,辛殿旗,那彥群;神經內分泌分化對前列腺癌細胞生長及雄激素受體表達影響的實驗研究[J];中華外科雜志;2004年23期

本文編號：2019389