CapG促进前列腺癌细胞增殖、迁移和侵袭的研究

发布时间：2018-04-30 09:54

本文选题：CapG + 前列腺癌　；参考：《南方医科大学》2017年博士论文

【摘要】：前列腺癌(prostate cancer,PCa)是全球男性发病率最高的恶性肿瘤。我国前列腺癌发病率低于欧美国家,但是我国前列腺癌患者的生存率却不容乐观,五年生存率仅为53.89%,大多数患者在初诊时就已经出现远处转移。因此,对具有高转移倾向的PCa患者的筛查和对已转移的晚期前列腺癌患者的治疗仍然是诸多挑战之一。本文通过研究前列腺癌中CapG的表达及其对前列腺癌细胞功能的影响揭示两者之间的关系。本研究首先通过生物信息学挖掘出与前列腺癌的发生和转移具有密切关系的差异基因CapG。对标本进行检测发现CapG在前列腺癌组织中阳性表达明显高于良性前列腺增生组织。与患者临床病理参数作相关性分析,发现CapG表达与患者年龄、吸烟史没有相关性,而与PSA、Gleason评分、T分级、淋巴结转移均具有显著的相关性。通过K-M生存曲线发现CapGp阳性会影响患者无生化复发生存率。我们通过qRT-PCR、Western blotting对三株前列腺癌细胞株LNCap、PC3、DU145和一株正常前列腺细胞株PrEC进行了检测,验证了 CapG在前列腺癌细胞株的表达与细胞的恶性程度及转移能力相关,且均显著性高于PrEC。构建 DU145-shCapG 和对照组 DU145-NC,LNCap-CapG 和对照组 LNCap-NC,通过qRT-PCR 和 Western blotting 验证 Du45-NC 和 LNCap-CapG 的 CapG 表达分别明显高于DU145-shCapG和LNCap-NC。功能实验结果显示:1、增殖实验:DU145-NC较DU145-shCapG的细胞生长显著加快;与LNCap-NC相比,LNCap-CapG细胞生长显著加快。提示CapG可促进前列腺癌细胞体外增殖能力。2、周期、凋亡实验:DU145-shCapG较DU145-NC细胞G1期显著增多,S期、G2期显著减少;LNCap-CapG较LNCap-NC细胞G1期显著减少,S期、G2期显著增多。DU 145-shCapG较DU 145-NC细胞凋亡数目显著增加,LNCap-CapG较LNCap-NC细胞凋亡数目显著降低。提示CapG通过促进细胞周期和抑制凋亡促进细胞的增殖。3、裸鼠皮下成瘤实验:DU145-NC成瘤体积显著大于DU145-ShCapG,提示CapG促进前列腺癌细胞体内的成瘤能力。4、划痕实验:DU145-NC相比,DU145-shCapG的迁移距离显著减少;LNCap-CapG的细胞迁移距离较LNCap-NC显著增加。表明CapG能够促进前列腺癌细胞株的体外迁移能力。5、Transwell实验:DU145-shCapG穿孔细胞数目显著低于DU145-NC;LNCap-CapG穿孔细胞数目显著高于LNCap-NC。说明CapG能够促进前列腺癌细胞体外侵袭能力。6、EMT:检测了前列腺癌细胞株中EMT相关分子标志物的mRNA以及蛋白质的表达,DU145-shCapG和LNCap-NC分别较对照组的 E-cadherin 表达升高,N-cadherin、vimentin、TWIST1 的表达降低。提示CapG通过促进EMT过程增强前列腺癌细胞株的侵袭、迁移能力。7、PI3K/AKT通路:检测前列腺癌细胞株中AKT和p-AKT的表达,DU145-NC较DU145-shCapG和LNCap-NC分别较对照组的p-AKT表达明显降低。两组细胞中ATK无显著性差异。推测CapG可能通过PI3K/AKT通路促进前列腺癌细胞株EMT过程。通过生物信息学及实验证实了与前列腺癌发生和转移密切相关的差异基因CapG。发现该基因与前列腺癌患者的临床病理参数存在相关性,并且与患者的无生化复发生存率相关。CapG能够促进前列腺癌细胞体内外增殖能力,同时促进前列腺癌细胞的体外侵袭和迁移能力。初步揭示了 CapG促进前列腺癌侵袭、迁移可能是通过PI3K/AKT通路介导的EMT途径实现的。
[Abstract]:Prostate cancer (PCa) is the highest incidence of male malignant tumor in the world. The incidence of prostate cancer in China is lower than that in European and American countries. However, the survival rate of prostate cancer patients in China is not optimistic and the five year survival rate is only 53.89%. Most patients have distant metastasis at first diagnosis. Therefore, the tendency of high metastasis is high. The screening of PCa patients and the treatment of advanced advanced prostate cancer patients are still one of the challenges. This paper reveals the relationship between the expression of CapG in prostate cancer and its effect on the function of prostate cancer cells. First, this study excavated the occurrence and metastasis of prostate cancer by bioinformatics. The positive expression of the closely related gene CapG. found that the positive expression of CapG in the prostate cancer tissues was significantly higher than that of the benign prostatic hyperplasia tissue. The correlation analysis with the clinicopathological parameters of the patients showed that the expression of CapG was not related to the age of the patients and the history of smoking, but with the PSA, the Gleason score, the T classification, and the lymph node metastasis. There was a significant correlation. Through the K-M survival curve, CapGp positive could affect the patient's survival rate without biochemical recurrence. We tested the PrEC of three prostate cancer cell lines, LNCap, PC3, DU145 and a normal prostate cell strain by qRT-PCR and Western blotting, and verified the expression of CapG in the prostate cancer cell line and the malignant cell line. The degree and ability to transfer were significantly higher than that of PrEC. in DU145-shCapG and control group DU145-NC, LNCap-CapG and control group LNCap-NC. The CapG expression of Du45-NC and LNCap-CapG through qRT-PCR and Western blotting was significantly higher than that of DU145-shCapG and functional experimental results, respectively: 1, proliferation experiment: Compared with DU145-shCapG, the growth of LNCap-CapG cells was significantly faster than that of LNCap-NC. It suggested that CapG could promote the proliferation of prostate cancer cells in vitro,.2, cycle and apoptosis experiment: DU145-shCapG was significantly increased in G1 phase of DU145-NC cells, S phase and G2 period decreased significantly, LNCap-CapG compared with LNCap-NC cells, period, period, period The number of.DU 145-shCapG increased significantly compared with the number of DU 145-NC cells, and the number of LNCap-CapG was significantly lower than that of LNCap-NC cells. It suggests that CapG can promote cell proliferation by promoting cell cycle and inhibiting apoptosis, and the tumor growth of nude mice is significantly larger than DU145-ShCapG, suggesting that CapG promotes prostate cancer cells. In vivo tumor formation ability.4, scratch test: compared with DU145-NC, the migration distance of DU145-shCapG decreased significantly, and the migration distance of LNCap-CapG cells increased significantly than LNCap-NC. It showed that CapG could promote the migration ability of prostate cancer cells in vitro.5, Transwell experiment: the number of DU145-shCapG perforated cells was significantly lower than DU145-NC; LNCap-CapG perforation. The number of cells was significantly higher than that of LNCap-NC. indicating that CapG could promote the invasion ability of prostate cancer cells in vitro.6. EMT: detected the mRNA and protein expression of EMT related molecular markers in the prostate cancer cell lines. DU145-shCapG and LNCap-NC were higher than those of the control group, and N-cadherin, vimentin, and TWIST1. It is suggested that CapG can enhance the invasion of the prostate cancer cell line by promoting the EMT process, the migration ability.7, the PI3K/AKT pathway: the expression of AKT and p-AKT in the prostate cancer cell line, DU145-NC is significantly lower than DU145-shCapG and LNCap-NC compared with the p-AKT expression in the control group. There is no significant difference between the two groups. The pathway promotes the EMT process of prostate cancer cell line. Through bioinformatics and experiments, the differential gene CapG., which is closely related to the occurrence and metastasis of prostate cancer, is found to be associated with the clinicopathological parameters of the prostate cancer patients and that.CapG can promote the prostate cancer cells with the survival rate of the patient's non biochemical recurrence. The ability to proliferate in vitro and in vivo promotes the invasion and migration of prostate cancer cells in vitro. It is preliminarily revealed that CapG promotes the invasion of prostate cancer, and migration may be achieved through the EMT pathway mediated by the PI3K/AKT pathway.

【学位授予单位】：南方医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R737.25

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