蛋白酶激活受体、IL-8在前列腺癌细胞株中表达的意义

发布时间：2018-06-15 23:08

本文选题：前列腺癌 + 蛋白酶激活受体　；参考：《昆明医科大学》2014年硕士论文

【摘要】：目的：观察蛋白酶激活受体PAR1,2,4与白细胞介素-8在不同前列腺癌细胞株的表达及作用,探索前列腺癌的分子病理机制。方法： 1、体外培养前列腺上皮细胞株HPrEpiC、雄激素依赖性前列腺癌细胞株LNCaP、雄激素非依赖性前列腺癌细胞株DU145、PC-3,并检测PAR1、PAR2,、PAR4,IL-8基因及蛋白的表达。 2、将细胞分组,分别以未加入激动剂0h及加入PAR1(20umol/L),PAR2(150umol/L), PAR4(300umol/L)激动剂后12h,24h,48h提取细胞,用MTT方法检测前列腺癌细胞增殖能力； 3、将细胞分组,分别加入PAR1(20umol/L), PAR2(150umol/L), PAR4(300umol/L)的激动剂,分未加入激动剂0h及加入激动剂后12h,24h,48h提取细胞,用Real-time PCR、Western blotting技术检测蛋白酶激活受体PAR1、PAR2、PAR4及IL-8的表达变化,探讨蛋白酶激活受体的激活与IL-8升高的密切关系。结果： 1、与前列腺上皮细胞相比,前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)中工L-8基因及蛋白表达明显升高(P0.05)；PAR1、PAR2,、PAR4基因表达升高(P0.05)； 2、MTT结果提示：三种前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)中,PAR1,2,4的激活均可加强前列腺癌细胞增殖能力(P0.05)；且PAR1、PAR2、PAR4激动效果无明显差异(P0.05)；同等条件下,三种前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)增殖能力无明显差异(P0.05)； 3、Real-time, western-blot检测结果提示：PAR1、PAR2、PAR4激动剂可以激活前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)及前列腺上皮细胞HPrEpiC中对应的PAR受体；被激活蛋白酶激活受体的前列腺癌细胞IL-8表达量明显升高,但前列腺上皮细胞HPrEpiC中IL-8变化不明显(P0.05)；雄激素非依赖性前列腺癌DU145、PC-3细胞中PAR和IL-8的表达要高于雄激素依赖性前列腺癌LNcap细胞。结论： 1、前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)和前列腺上皮细胞HPrEpiC中均有PAR1、PAR2、PAR4及IL-8基因及蛋白的表达；前列腺癌细胞中PAR及IL-8基因及蛋白表达要高于前列腺上皮细胞；非激素依赖性前列腺癌细胞中PAR及IL-8基因及蛋白表达要高于激素依赖性前列腺癌； 2、IL-8的含量与前列腺癌(LNcap细胞、PC-3细胞、DU145细胞)细胞的增殖能力有正相关性。 3、蛋白酶激活受体的升高可激活前列腺癌细胞(LNcap细胞、PC-3细胞、DU145细胞)的增殖能力,同等增殖能力下,非激素依赖性前列腺癌(PC-3细胞、DU145细胞)中IL-8基因及蛋白的表达更高。 4、蛋白酶激活受体可能会成为前列腺癌药物治疗的新靶点。
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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