本文選題：前列腺腫瘤 + miR-19b-1-5p��； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：研究目的:前列腺癌(prostate cancer,PCa)主要發(fā)生于老年人群。數(shù)年前,我國前列腺癌的發(fā)病率還處在一個(gè)很低的程度,但隨著人們生活水準(zhǔn)的逐漸西式化以及診斷程度的不斷上升,我國前列腺癌的檢出率和發(fā)病率開始連年增高,已然成為危及中老年男性生命的重要疾病。本研究基于微小RNA(miRNA,miR)對(duì)前列腺癌存在抑制或促進(jìn)的作用而產(chǎn)生,旨在探討miR-19b-1-5p對(duì)人前列腺癌細(xì)胞生物學(xué)行為的影響,闡明miR-19b-1-5p在前列腺癌中的作用以及發(fā)生發(fā)展的主要機(jī)制。研究方法:收集臨床標(biāo)本,劃分前列腺癌組和良性前列腺組織組,各組標(biāo)本分別為77例和68例。實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)技術(shù)(Real-time PCR)檢測各組織中miR-19b-1-5p的表達(dá)情況,分析mi R-19b-1-5p的相對(duì)表達(dá)量與前列腺癌Gleason評(píng)分和臨床病理特征之間的關(guān)系。Real-time PCR檢測不同前列腺癌細(xì)胞系和正常前列腺上皮細(xì)胞中miR-19b-1-5p的表達(dá)情況。將miR-19b-1-5p的模擬物(mimic)、抑制物(inhibitor)經(jīng)化學(xué)轉(zhuǎn)染試劑分別轉(zhuǎn)染進(jìn)人前列腺癌細(xì)胞系中。細(xì)胞計(jì)數(shù)(Cell Counting Kit-8,CCK-8)試劑盒檢測細(xì)胞的增殖能力變化,Transwell小室檢測細(xì)胞的侵襲和轉(zhuǎn)移能力,劃痕實(shí)驗(yàn)檢測細(xì)胞遷移能力,流式細(xì)胞儀分析細(xì)胞凋亡能力,以此探究miR-19b-1-5p對(duì)前列腺癌細(xì)胞生物學(xué)行為的影響及其作用。應(yīng)用靶基因預(yù)測軟件miRanda、TargetScan、PicTar等預(yù)測miR-19b-1-5p的潛在靶基因。Real-time PCR檢測miR-19b-1-5p與靶基因mRNA的相對(duì)表達(dá)量,分析二者之間的相互關(guān)系,Western blot驗(yàn)證靶基因的表達(dá)變化。結(jié)果:mi R-19b-1-5p在良性前列腺組織內(nèi)高表達(dá),而在前列腺癌組織中表達(dá)量較低,兩組比較差異有顯著性(t=24.87,P0.05),miR-19b-1-5p的相對(duì)表達(dá)量與腫瘤的Gleason評(píng)分、臨床分期間有關(guān)。前列腺癌細(xì)胞系PC-3、DU145、LNCap、C4-2同正常前列腺上皮細(xì)胞RWPE-1比較發(fā)現(xiàn),miR-19b-1-5p在前列腺癌細(xì)胞系中表達(dá)量較低,RWPE-1中表達(dá)量較高,而在DU145、PC-3中表達(dá)量最低。CCK-8實(shí)驗(yàn)表明轉(zhuǎn)染miR-19b-1-5p-mimic組的細(xì)胞增殖率顯著低于miR-19b-1-5p-inhibitor組。Transwell實(shí)驗(yàn)中,轉(zhuǎn)染mi R-19b-1-5p-mimic組的細(xì)胞通過含基質(zhì)膠的Transwell小室數(shù)量明顯少于miR-19b-1-5p-inhibitor組。劃痕實(shí)驗(yàn)中,轉(zhuǎn)染miR-19b-1-5p-mimic組的細(xì)胞閉合率明顯小于miR-19b-1-5p-inhibitor組。流式分析實(shí)驗(yàn)顯示轉(zhuǎn)染miR-19b-1-5p-mimic組的細(xì)胞凋亡率增加。靶基因軟件預(yù)測CXCR6為mi R-19b-1-5p的下游靶基因,Real-time PCR結(jié)果顯示miR-19b-1-5p與CXCR6存在負(fù)相關(guān),Western blot實(shí)驗(yàn)也證實(shí)轉(zhuǎn)染miR-19b-1-5p-mimic后CXCR6的表達(dá)水平受到抑制。結(jié)論及意義:miR-19b-1-5p在前列腺癌組織內(nèi)低表達(dá),起抑癌基因的作用。隨著Gleason評(píng)分的增高、臨床分期級(jí)別增高而表達(dá)量減少。上調(diào)miR-19b-1-5p的表達(dá)可明顯抑制前列腺癌細(xì)胞株的增殖、侵襲和轉(zhuǎn)移能力,促進(jìn)其凋亡,可能是通過抑制下游靶基因CXCR6的表達(dá)實(shí)現(xiàn)。
[Abstract]:Objective: prostate cancer (PCA) mainly occurs in the elderly.A few years ago, the incidence of prostate cancer in China was still at a very low level. However, with the gradual westernization of people's standard of living and the rising diagnostic level, the detection rate and incidence of prostate cancer in China began to increase year after year.It has become an important disease that endangers the life of middle-aged and aged men.The aim of this study was to investigate the effect of miR-19b-1-5p on the biological behavior of human prostate cancer cells and to elucidate the role of miR-19b-1-5p in prostate cancer and the main mechanism of its development.Methods: clinical specimens were collected and divided into prostate cancer group (77 cases) and benign prostate tissue group (68 cases).Real-time PCR was used to detect the expression of miR-19b-1-5p in tissues.To analyze the relationship between the relative expression of mi R-19b-1-5p and Gleason score and clinicopathological features of prostate cancer. Real-time PCR was used to detect the expression of miR-19b-1-5p in different prostate cancer cell lines and normal prostate epithelial cells.The mimic of miR-19b-1-5p was transfected into human prostate cancer cell line by chemical transfection reagent.Cell Counting Kit-8 CCK-8 kit was used to detect cell proliferation ability. Cell invasion and metastasis were detected by Transwell chamber, migration ability was detected by scratch assay, and apoptosis was analyzed by flow cytometry.To investigate the effect of miR-19b-1-5p on the biological behavior of prostate cancer cells.The relative expression of miR-19b-1-5p and target gene mRNA was detected by using the target gene prediction software miRanda Target Scann PicTar to predict the potential target gene. Real-time PCR was used to detect the relative expression of miR-19b-1-5p and target gene mRNA, and the correlation between them was analyzed. Western blot was used to verify the change of target gene expression.Results there was a significant difference between the two groups in the high expression of R-19b-1-5p in the benign prostate tissue and the low expression in the prostate cancer tissue. The relative expression of the two groups was correlated with the Gleason score of the tumor and the clinical stage.Compared with normal prostatic epithelial cells (RWPE-1), prostate cancer cell line PC-3DU145LNCapC4-2 was found to have a higher expression level of miR-19b-1-5p in prostate cancer cell line than that in prostate cancer cell line RWPE-1.CCK-8 assay showed that the cell proliferation rate of miR-19b-1-5p-mimic group was significantly lower than that of miR-19b-1-5p-inhibitor group. The number of Transwell cells transfected with mi R-19b-1-5p-mimic group was significantly lower than that of miR-19b-1-5p-inhibitor group.In scratch test, the cell closure rate of miR-19b-1-5p-mimic transfected group was significantly lower than that of miR-19b-1-5p-inhibitor group.Flow cytometry showed that the apoptosis rate of miR-19b-1-5p-mimic transfected group was increased.Real-time PCR of CXCR6 as a downstream target gene of mi R-19b-1-5p was predicted by target gene software. The results showed that there was a negative correlation between miR-19b-1-5p and CXCR6. Western blot experiment also confirmed that the expression of CXCR6 was inhibited after transfection of miR-19b-1-5p-mimic.Conclusion the low expression of 1: miR-19b-1-5p in prostate cancer may play an important role in tumor suppressor gene.With the increase of Gleason score, the clinical stage grade increased and the expression decreased.Upregulating the expression of miR-19b-1-5p can obviously inhibit the ability of proliferation, invasion and metastasis of prostate cancer cell line and promote its apoptosis, which may be realized by inhibiting the expression of downstream target gene CXCR6.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.25

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相關(guān)期刊論文 前2條

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本文編號(hào)：1749026