本文選題：前列腺癌 + 蛋白酶激活受體�。� 參考：《昆明醫(yī)科大學》2014年碩士論文

【摘要】：目的： 觀察蛋白酶激活受體PAR1,2,4與白細胞介素-8在不同前列腺癌細胞株的表達及作用,探索前列腺癌的分子病理機制。方法： 1、體外培養(yǎng)前列腺上皮細胞株HPrEpiC、雄激素依賴性前列腺癌細胞株LNCaP、雄激素非依賴性前列腺癌細胞株DU145、PC-3,并檢測PAR1、PAR2,、PAR4,IL-8基因及蛋白的表達。 2、將細胞分組,分別以未加入激動劑0h及加入PAR1(20umol/L),PAR2(150umol/L), PAR4(300umol/L)激動劑后12h,24h,48h提取細胞,用MTT方法檢測前列腺癌細胞增殖能力； 3、將細胞分組,分別加入PAR1(20umol/L), PAR2(150umol/L), PAR4(300umol/L)的激動劑,分未加入激動劑0h及加入激動劑后12h,24h,48h提取細胞,用Real-time PCR、Western blotting技術(shù)檢測蛋白酶激活受體PAR1、PAR2、PAR4及IL-8的表達變化,探討蛋白酶激活受體的激活與IL-8升高的密切關(guān)系。 結(jié)果： 1、與前列腺上皮細胞相比,前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)中工L-8基因及蛋白表達明顯升高(P0.05)；PAR1、PAR2,、PAR4基因表達升高(P0.05)； 2、MTT結(jié)果提示：三種前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)中,PAR1,2,4的激活均可加強前列腺癌細胞增殖能力(P0.05)；且PAR1、PAR2、PAR4激動效果無明顯差異(P0.05)；同等條件下,三種前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)增殖能力無明顯差異(P0.05)； 3、Real-time, western-blot檢測結(jié)果提示：PAR1、PAR2、PAR4激動劑可以激活前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)及前列腺上皮細胞HPrEpiC中對應(yīng)的PAR受體；被激活蛋白酶激活受體的前列腺癌細胞IL-8表達量明顯升高,但前列腺上皮細胞HPrEpiC中IL-8變化不明顯(P0.05)；雄激素非依賴性前列腺癌DU145、PC-3細胞中PAR和IL-8的表達要高于雄激素依賴性前列腺癌LNcap細胞。 結(jié)論： 1、前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)和前列腺上皮細胞HPrEpiC中均有PAR1、PAR2、PAR4及IL-8基因及蛋白的表達；前列腺癌細胞中PAR及IL-8基因及蛋白表達要高于前列腺上皮細胞；非激素依賴性前列腺癌細胞中PAR及IL-8基因及蛋白表達要高于激素依賴性前列腺癌； 2、IL-8的含量與前列腺癌(LNcap細胞、PC-3細胞、DU145細胞)細胞的增殖能力有正相關(guān)性。 3、蛋白酶激活受體的升高可激活前列腺癌細胞(LNcap細胞、PC-3細胞、DU145細胞)的增殖能力,同等增殖能力下,非激素依賴性前列腺癌(PC-3細胞、DU145細胞)中IL-8基因及蛋白的表達更高。 4、蛋白酶激活受體可能會成為前列腺癌藥物治療的新靶點。
[Abstract]:Aim: to investigate the expression and role of protease activated receptor PAR1O2O4 and interleukin-8 (IL-8) in different prostate cancer cell lines, and to explore the molecular pathological mechanism of prostate cancer. Methods: 1. Prostatic epithelial cell line HPrEpiCin, androgen dependent prostate cancer cell line LNCaP, androgen independent prostate cancer cell line DU145HPC-3 were cultured in vitro. The cells were extracted from the prostate cancer cells at 12 h and 24 h after the agonist was added for 0 h, and the PAR1 + 20 umol / L + PAR2150 umolr / L, and the PAR4 + 300umol / L) agonist for 48 h, respectively, and the proliferation of prostate cancer cells was measured by MTT assay. 3. The cells were divided into two agonists, PAR1 20 umoll / L, PAR2 + 150 umoll / L, PAR4R4300umolrL, respectively, and the cells were divided into two agonists, PAR120umolrL, PAR2OL, PAR4300umolrL, respectively. The cells were extracted at 24 h and 48 h after no agonist was added and the expression of PAR4 and IL-8 in the protease activated receptor (PAR1), PAR2, PAR4 and IL-8 were detected by Real-time PCR blotting. The close relationship between the activation of protease activated receptor and the increase of IL-8 was discussed. Results: 1, compared with the prostatic epithelial cells, the expression of L-8 gene and protein in the prostate cancer cell line LNcap / PC-3 cell line (DU145) increased significantly (P 0.05) and the expression of the PAR2 / PAR4 gene increased significantly (P 0.05). The results of MTT indicated that the activation of PAR1O2O4 in three kinds of prostate cancer cell line (LNcap) and PC-3 cell line (DU145) could enhance the proliferative ability of prostate cancer cells (P0.05), and there was no significant difference in the stimulative effect of PAR1, PAR2, PAR4, and under the same conditions, there was no significant difference in the activation effect of PAR1, PAR2, PAR4 and P0. 05 in the same condition. There was no significant difference in the proliferative ability of three kinds of prostate cancer cells (LNcap cell line, PC-3 cell line, DU145 cell line). 3Real-time, western-blot results showed that the PAR4 agonist could activate the par receptor in the prostate cancer cell line (LNcap) and in the prostate epithelial cell (HPrEpiC), and in the prostatic epithelial cell (HPrEpiC). The results showed that the PAR4 agonist could activate the receptor in the prostate cancer cell line (LNcap) and in the prostate epithelial cell line (HPrEpiC). The expression of IL-8 in prostate cancer cells activated by protease-activated receptors was significantly increased, but the expression of IL-8 in HPREpiC cells did not change significantly (P 0.05). The expression of par and IL-8 in androgen independent prostate cancer DU145 PC-3 cells was higher than that in androgen dependent prostate cancer LNcap cells. Conclusion: 1, the expression of PAR1PAR2PAR4 and IL-8 gene and protein in prostate cancer cell line (LNcap) and prostatic epithelial cell (HPREpiC) and the expression of par and IL-8 gene and protein in prostate cancer cell were higher than those in prostate epithelial cell. The expression of par and IL-8 gene and protein in hormone independent prostate cancer cells was higher than that in hormone dependent prostate cancer cells. 2There was a positive correlation between the content of IL-8 and the proliferative ability of human prostate cancer cell line DU145. 3. The increase of protease activated receptor could activate the proliferation of prostate cancer cell line LNcap and PC-3 cell line DU145. The expression of IL-8 gene and protein in PC-3 cell line was higher than that in DU145 cell line. 4. Protease activated receptor may become a new target of drug therapy for prostate cancer.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.25

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