本文選題：CapG + 前列腺癌��； 參考：《南方醫(yī)科大學(xué)》2017年博士論文

【摘要】：前列腺癌(prostate cancer,PCa)是全球男性發(fā)病率最高的惡性腫瘤。我國(guó)前列腺癌發(fā)病率低于歐美國(guó)家,但是我國(guó)前列腺癌患者的生存率卻不容樂(lè)觀,五年生存率僅為53.89%,大多數(shù)患者在初診時(shí)就已經(jīng)出現(xiàn)遠(yuǎn)處轉(zhuǎn)移。因此,對(duì)具有高轉(zhuǎn)移傾向的PCa患者的篩查和對(duì)已轉(zhuǎn)移的晚期前列腺癌患者的治療仍然是諸多挑戰(zhàn)之一。本文通過(guò)研究前列腺癌中CapG的表達(dá)及其對(duì)前列腺癌細(xì)胞功能的影響揭示兩者之間的關(guān)系。本研究首先通過(guò)生物信息學(xué)挖掘出與前列腺癌的發(fā)生和轉(zhuǎn)移具有密切關(guān)系的差異基因CapG。對(duì)標(biāo)本進(jìn)行檢測(cè)發(fā)現(xiàn)CapG在前列腺癌組織中陽(yáng)性表達(dá)明顯高于良性前列腺增生組織。與患者臨床病理參數(shù)作相關(guān)性分析,發(fā)現(xiàn)CapG表達(dá)與患者年齡、吸煙史沒(méi)有相關(guān)性,而與PSA、Gleason評(píng)分、T分級(jí)、淋巴結(jié)轉(zhuǎn)移均具有顯著的相關(guān)性。通過(guò)K-M生存曲線發(fā)現(xiàn)CapGp陽(yáng)性會(huì)影響患者無(wú)生化復(fù)發(fā)生存率。我們通過(guò)qRT-PCR、Western blotting對(duì)三株前列腺癌細(xì)胞株LNCap、PC3、DU145和一株正常前列腺細(xì)胞株P(guān)rEC進(jìn)行了檢測(cè),驗(yàn)證了 CapG在前列腺癌細(xì)胞株的表達(dá)與細(xì)胞的惡性程度及轉(zhuǎn)移能力相關(guān),且均顯著性高于PrEC。構(gòu)建 DU145-shCapG 和對(duì)照組 DU145-NC,LNCap-CapG 和對(duì)照組 LNCap-NC,通過(guò)qRT-PCR 和 Western blotting 驗(yàn)證 Du45-NC 和 LNCap-CapG 的 CapG 表達(dá)分別明顯高于DU145-shCapG和LNCap-NC。功能實(shí)驗(yàn)結(jié)果顯示:1、增殖實(shí)驗(yàn):DU145-NC較DU145-shCapG的細(xì)胞生長(zhǎng)顯著加快;與LNCap-NC相比,LNCap-CapG細(xì)胞生長(zhǎng)顯著加快。提示CapG可促進(jìn)前列腺癌細(xì)胞體外增殖能力。2、周期、凋亡實(shí)驗(yàn):DU145-shCapG較DU145-NC細(xì)胞G1期顯著增多,S期、G2期顯著減少;LNCap-CapG較LNCap-NC細(xì)胞G1期顯著減少,S期、G2期顯著增多。DU 145-shCapG較DU 145-NC細(xì)胞凋亡數(shù)目顯著增加,LNCap-CapG較LNCap-NC細(xì)胞凋亡數(shù)目顯著降低。提示CapG通過(guò)促進(jìn)細(xì)胞周期和抑制凋亡促進(jìn)細(xì)胞的增殖。3、裸鼠皮下成瘤實(shí)驗(yàn):DU145-NC成瘤體積顯著大于DU145-ShCapG,提示CapG促進(jìn)前列腺癌細(xì)胞體內(nèi)的成瘤能力。4、劃痕實(shí)驗(yàn):DU145-NC相比,DU145-shCapG的遷移距離顯著減少;LNCap-CapG的細(xì)胞遷移距離較LNCap-NC顯著增加。表明CapG能夠促進(jìn)前列腺癌細(xì)胞株的體外遷移能力。5、Transwell實(shí)驗(yàn):DU145-shCapG穿孔細(xì)胞數(shù)目顯著低于DU145-NC;LNCap-CapG穿孔細(xì)胞數(shù)目顯著高于LNCap-NC。說(shuō)明CapG能夠促進(jìn)前列腺癌細(xì)胞體外侵襲能力。6、EMT:檢測(cè)了前列腺癌細(xì)胞株中EMT相關(guān)分子標(biāo)志物的mRNA以及蛋白質(zhì)的表達(dá),DU145-shCapG和LNCap-NC分別較對(duì)照組的 E-cadherin 表達(dá)升高,N-cadherin、vimentin、TWIST1 的表達(dá)降低。提示CapG通過(guò)促進(jìn)EMT過(guò)程增強(qiáng)前列腺癌細(xì)胞株的侵襲、遷移能力。7、PI3K/AKT通路:檢測(cè)前列腺癌細(xì)胞株中AKT和p-AKT的表達(dá),DU145-NC較DU145-shCapG和LNCap-NC分別較對(duì)照組的p-AKT表達(dá)明顯降低。兩組細(xì)胞中ATK無(wú)顯著性差異。推測(cè)CapG可能通過(guò)PI3K/AKT通路促進(jìn)前列腺癌細(xì)胞株EMT過(guò)程。通過(guò)生物信息學(xué)及實(shí)驗(yàn)證實(shí)了與前列腺癌發(fā)生和轉(zhuǎn)移密切相關(guān)的差異基因CapG。發(fā)現(xiàn)該基因與前列腺癌患者的臨床病理參數(shù)存在相關(guān)性,并且與患者的無(wú)生化復(fù)發(fā)生存率相關(guān)。CapG能夠促進(jìn)前列腺癌細(xì)胞體內(nèi)外增殖能力,同時(shí)促進(jìn)前列腺癌細(xì)胞的體外侵襲和遷移能力。初步揭示了 CapG促進(jìn)前列腺癌侵襲、遷移可能是通過(guò)PI3K/AKT通路介導(dǎo)的EMT途徑實(shí)現(xiàn)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.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R737.25

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1 木海琦;CapG促進(jìn)前列腺癌細(xì)胞增殖、遷移和侵襲的研究[D];南方醫(yī)科大學(xué);2017年

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本文編號(hào)：1824085