本文選題：前列腺癌 + 臨床病理特征��； 參考：《新疆醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:研究前列腺癌患者組織中基因的甲基化水平、免疫細(xì)胞表面Tim-3表達(dá)水平與前列腺癌臨床病理特征的關(guān)系,探討甲基化和Tim-3在前列腺癌發(fā)生、侵襲和轉(zhuǎn)移中的作用。方法:1)選取PITX2、WNT5a、SPARC、EPB41L3、TPM4等基因作為目的基因,然后通過甲基化特異性PCR方法分別檢測(cè)35例良性前列腺增生組織中甲基化水平,接著再對(duì)157例臨床DNA樣本進(jìn)行篩選,分析甲基化水平與前列腺癌臨床病理特征的關(guān)系;2)利用流式細(xì)胞分析法檢測(cè)35例良性前列腺增生癥患者和157例前列腺癌患者血清中免疫細(xì)胞Tim-3表達(dá)水平,分析血清中Tim-3表達(dá)水平與前列腺癌臨床病理特征的關(guān)系;采用免疫組織化學(xué)方法檢測(cè)組織中Tim-3在35例良性前列腺增生癥和157例前列腺癌標(biāo)本中的表達(dá)水平,統(tǒng)計(jì)分析組織中Tim-3表達(dá)水平與前列腺癌臨床病理特征的關(guān)系;3)采用western blot法檢測(cè)轉(zhuǎn)移潛能前列腺癌LNCa P細(xì)胞中Tim-3的表達(dá)水平,分析其表達(dá)水平與前列腺癌侵襲轉(zhuǎn)移能力;采用Si RNA干擾前列腺癌LNCa P細(xì)胞中Tim-3的表達(dá),MTT檢測(cè)細(xì)胞增殖,Transwell小室實(shí)驗(yàn)檢測(cè)前列腺癌LNCa P細(xì)胞侵襲能力變化。結(jié)果:1)前列腺癌患者組織中PITX2,WNT5a,SPARC,EPB41L3,和TPM4甲基化水平明顯升高,提示這些基因甲基化水平與前列腺癌發(fā)生發(fā)展有一定的關(guān)系。根據(jù)不同PSA水平結(jié)果顯示PSA10ng/ml、10~20ng/ml、20ng/ml組PITX2甲基化率分別為25.10%、28.64%及35.82%,不同PSA組中PITX2及WNT5a基因甲基化水平比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05);Glenson評(píng)分≤7分組和≥8分組,PITX2基因甲基化率分別為26.31%、33.16%,兩組間比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);根據(jù)不同TNM分期分組為≤T2期、Tx N1期及M1期組,PITX2基因甲基化率為23.13%、26.64%、38.37%,組間比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)。多因素分析發(fā)現(xiàn)PITX2甲基化水平與前列腺癌患者的關(guān)系最密切。2)前列腺癌患者外周血中CD4+T、CD8+T細(xì)胞表面Tim-3表達(dá)水平分別為(4.77±3.56)%,(5.90±4.91)%,良性前列腺增生的表達(dá)量為(0.96±0.54)%,(0.73±0.62)%,差異有統(tǒng)計(jì)學(xué)意義(P0.05);前列腺增生和前列腺癌組中Tim-3免疫組化表達(dá)水平評(píng)分比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);前列腺癌免疫組化染色Tim-3表達(dá)水平據(jù)不同PSA水平結(jié)果顯示,PSA10ng/ml、10~20ng/ml、20ng/ml組Tim-3免疫組化評(píng)分分別為3.92±0.75分、7.63±0.81分及10.37±0.35分,差異有統(tǒng)計(jì)學(xué)意義(P0.05);根據(jù)不同Glenson評(píng)分,分為≤7分組和≥8分組,Tim-3免疫組化評(píng)分為11.02±0.96分,5.62±0.69分,差異有統(tǒng)計(jì)學(xué)意義(P0.05);根據(jù)不同TNM分期分組為≤T2期、Tx N1期及M1期組,Tim-3免疫組化評(píng)分為4.27±0.38,7.34±0.71,10.31±0.82,差異有統(tǒng)計(jì)學(xué)意義(P0.05)3)Tim-3對(duì)人前列腺癌細(xì)胞增殖、侵襲能力結(jié)果提示,對(duì)照組和過表達(dá)組和干擾組三組細(xì)胞:過表達(dá)組的Tim-3的表達(dá)水平顯著高于對(duì)照組,而干擾組Tim-3表達(dá)顯著低于對(duì)照組(P0.05)。結(jié)論:1)前列腺癌患者組織PITX2基因甲基化水平高于良性前列腺增生;PITX2甲基化水平與PSA水平、Gleason評(píng)分及TNM分期有關(guān),提示PITX2甲基化水平與前列腺癌的存在密切關(guān)系;2)前列腺癌患者血清及組織中Tim-3表達(dá)水平均高于良性前列腺增生;Tim-3表達(dá)水平與PSA水平、Gleason評(píng)分及TNM分期有關(guān),提示Tim-3表達(dá)水平與前列腺癌存在密切的關(guān)系;3)Tim-3表達(dá)下調(diào)能抑制前列腺癌細(xì)胞的生長,Tim-3是前列腺癌的重要調(diào)節(jié)因子;Tim-3表達(dá)水平與前列腺癌侵襲、轉(zhuǎn)移能力有關(guān),可作為前列腺癌侵襲、轉(zhuǎn)移的分子標(biāo)記物;下調(diào)Tim-3能抑制前列腺癌細(xì)胞的體外侵襲和遷移能力。
[Abstract]:Objective: To study the methylation level of gene in the tissues of prostate cancer patients, the relationship between the expression level of Tim-3 and the clinicopathological features of prostate cancer, and to explore the role of methylation and Tim-3 in the occurrence, invasion and metastasis of prostate cancer. Methods: 1) select PITX2, WNT5a, SPARC, EPB41L3, TPM4 and other genes as the target genes, and then pass through the gene as the target gene. Methylation specific PCR method was used to detect the methylation level in 35 cases of benign prostatic hyperplasia, and then 157 clinical DNA samples were screened to analyze the relationship between the methylation level and the clinicopathological features of the prostate cancer. 2) 35 cases of benign prostatic hyperplasia and 157 cases of prostate cancer were detected by flow cytometry. The expression level of immune cell Tim-3 in serum, the relationship between the expression of Tim-3 in serum and the clinicopathological features of prostate cancer, and the expression level of Tim-3 in 35 specimens of benign prostatic hyperplasia and 157 cases of prostate cancer by immunohistochemical method, and the level of Tim-3 expression in the analysis tissue and prostate cancer The relationship between the clinicopathological features; 3) the expression level of Tim-3 in the LNCa P cells of metastatic potential prostate cancer was detected by Western blot method, and the expression level of the prostate cancer and the invasion and metastasis of prostate cancer were analyzed. Si RNA interfered with the expression of Tim-3 in the LNCa P cells of the prostate cancer, and the MTT detected the proliferation of the cell, and the Transwell chamber test detected the prostate cancer. Results: 1) 1) the levels of PITX2, WNT5a, SPARC, EPB41L3, and TPM4 methylation in the tissues of the patients with prostate cancer were significantly elevated, suggesting that the levels of methylation of these genes were related to the development of prostate cancer. According to the results of PSA, the PITX2 methylation rate of PSA10ng /ml, 10~20ng/ml and 20ng/ml groups was 25.10%, respectively. 28.64% and 35.82%, the difference of PITX2 and WNT5a gene methylation levels in different PSA groups was statistically significant (P0.05); Glenson score was less than 7 groups and 8 groups, and the methylation rate of PITX2 gene was 26.31%, 33.16%, respectively, and two groups were statistically significant (P0.05); according to the different TNM stages, the group was less than T2, Tx N1 phase and M1 period group, The rate of gene methylation was 23.13%, 26.64%, 38.37%. The difference between the groups was statistically significant (P0.05). The multifactor analysis found that the level of PITX2 methylation was most closely related to the prostate cancer patients. The expression level of Tim-3 in the peripheral blood of the prostate cancer patients was (4.77 + 3.56)%, (5.90 + 4.91)%, and benign prostatic hyperplasia, respectively, in the peripheral blood of the prostate cancer patients. The amount of expression was (0.96 + 0.54)%, (0.73 + 0.62)%, and the difference was statistically significant (P0.05). The difference of Tim-3 immunohistochemical expression level in prostatic hyperplasia and prostate cancer group was statistically significant (P0.05), and the expression level of Tim-3 in prostate cancer immunohistochemical staining showed that PSA10ng/ml, 10~20ng/ml, 20ng/ml group Tim-3, according to the results of different PSA levels. The score of immunohistochemistry was 3.92 + 0.75, 7.63 + 0.81 and 10.37 + 0.35. The difference was statistically significant (P0.05). According to the different Glenson scores, it was divided into less than 7 groups and more than 8 groups. The Tim-3 immunohistochemical score was 11.02 + 0.96, 5.62 + 0.69, and the difference was statistically significant (P0.05). According to the different TNM stages, the group was less than T2, Tx N1 period and M1 Stage group, Tim-3 immunohistochemical score was 4.27 + 0.38,7.34 + 0.71,10.31 + 0.82, the difference was statistically significant (P0.05) 3) Tim-3 on human prostate cancer cell proliferation, invasive ability results suggested that the control group and overexpression group and interference group three groups of cells: the expression level of Tim-3 in the over expression group was significantly higher than the control group, while the Tim-3 expression in the interference group was significantly lower. In the control group (P0.05). Conclusion: 1) the methylation level of PITX2 gene in the tissues of the prostate cancer patients is higher than that of the benign prostatic hyperplasia; the level of PITX2 methylation is related to the level of PSA, the Gleason score and the TNM staging, suggesting that the level of PITX2 methylation is closely related to the existence of prostate cancer; and 2) the level of Tim-3 expression in the serum and tissue of the prostate cancer patients is higher than that of the prostate cancer patients. Benign prostatic hyperplasia; Tim-3 expression level is related to PSA level, Gleason score and TNM staging, suggesting that the expression level of Tim-3 is closely related to prostate cancer; 3) down regulation of Tim-3 expression can inhibit the growth of prostate cancer cells, Tim-3 is an important regulator of prostate cancer; the level of Tim-3 expression is related to the invasion and metastasis of prostate cancer. It can be used as a molecular marker for invasion and metastasis of prostate cancer, and down-regulation of Tim-3 can inhibit the invasion and migration ability of prostate cancer cells in vitro.

【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R737.25

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