發(fā)布時(shí)間：2018-03-13 04:34

  本文選題：microRNA-187　切入點(diǎn)：CD276　出處：《江蘇大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的探討microRNA-187(miR-187)在胃癌中的表達(dá)及對胃癌增殖、侵襲和遷移的影響及其機(jī)制。方法收集24對人胃癌組織和癌旁正常組織,檢測miR-187在胃癌組織和癌旁正常組織的表達(dá),探討其與各臨床病理指標(biāo)之間關(guān)系。檢測miR-187相關(guān)基因CD276在胃癌組織組織中的表達(dá)并分析兩者的相關(guān)性。分別抑制人胃腺癌細(xì)胞SGC-7901中miR-187和CD276的表達(dá);用CCK-8法檢測SGC-7901細(xì)胞中miR-187和CD276表達(dá)發(fā)生改變后對細(xì)胞增殖的影響;用Transwell法檢測miR-187和CD276表達(dá)發(fā)生改變后SGC-7901細(xì)胞侵襲性及遷移性的改變;用Western blot檢測SGC-7901細(xì)胞miR-187和CD276表達(dá)發(fā)生改變后基質(zhì)金屬蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表達(dá)改變。結(jié)果1.實(shí)時(shí)熒光定量PCR結(jié)果顯示miR-187在胃癌組織中表達(dá)升高,而在癌旁組織中低表達(dá)(P0.001)。Western blot檢測證實(shí)CD276在胃癌組織中表達(dá)降低,而在癌旁組織中高表達(dá)。miR-187的表達(dá)強(qiáng)度與胃癌患者淋巴結(jié)轉(zhuǎn)移個(gè)數(shù)(P=0.046)、TNM分期(P=0.015)及Cerb B-2(P=0.013)的表達(dá)相關(guān),而與患者的年齡、性別、腫瘤的部位、腫瘤大小、浸潤深度及Ki67的表達(dá)無關(guān)(P均0.05)。miR-187在胃癌淋巴結(jié)轉(zhuǎn)移≥3枚、III期和IV期及Cerb B-2陽性的胃癌組織中表達(dá)明顯強(qiáng)于淋巴結(jié)轉(zhuǎn)移小于3枚、I期和II期及Cerb B-2陰性的胃癌組織。CD276的表達(dá)強(qiáng)度與淋巴結(jié)轉(zhuǎn)移個(gè)數(shù)(P=0.044)、浸潤深度(P=0.042)、TNM分期(P=0.001)及Cerb B-2(P0.001)的表達(dá)相關(guān),而與患者的年齡、性別、腫瘤的部位、腫瘤大小及Ki67的表達(dá)無關(guān)(P均0.05)。CD276在腫瘤淋巴結(jié)轉(zhuǎn)移≥3枚、T3~T4、III期和IV期及Cerb B-2陽性的胃癌組織中表達(dá)明顯低于淋巴結(jié)轉(zhuǎn)移小于3枚、T1~T2、I期和II期及Cerb B-2陰性的胃癌組織。2.與癌旁正常組織比較,miR-187在I期、II期、III期和IV期胃癌組織表達(dá)進(jìn)行性升高,分別是正常組織的20.3倍、3.5倍、16.3倍和11.8倍(P0.001);CD276在I期、II期、III期和IV期胃癌組織表達(dá)降低,分別是正常組織的0.68倍、0.71倍、0.67倍和0.61倍(P0.01);miR-187在胃腺癌細(xì)胞SGC中的表達(dá)是胃癌粘膜上皮細(xì)胞GES的5.5倍(P0.001)。3.胃癌組織中miR-187表達(dá)與CD276表達(dá)相關(guān)系數(shù)R2值為0.840,表示在此胃癌標(biāo)本中miR-187表達(dá)量可以解釋CD276表達(dá)量變異性的76.63%,另外23.37%的變異不能用miR-187表達(dá)量解釋。4.與對照組(miR-187 inhibitor-control組)比較,miR-187表達(dá)抑制組(miR-187inhibitor)SGC增殖活性較對照組降低(60.67±11.02)%(P=0.003)、侵襲性較對照組降低(60.67±11.02)%(P=0.001)和遷移能力較對照組降低(66.33±10.6)%(P0.001);miR-187 inhibitor組CD276表達(dá)增加(1.047±0.06506)倍(P0.001)和MMP-9表達(dá)降低(0.2667±0.06028)倍(P0.001)。5.與對照組(CD276-siRNA-control組)比較,CD276表達(dá)干擾組(CD276-siRNA)SGC增殖活性為對照組的(1.823±0.2577)倍(P=0.002)、侵襲性為對照組的(1.843±0.3296)倍(P0.001)與遷移性為對照組的(1.66±0.1453)倍(P0.001);CD276-siRNA組CD276表達(dá)降低(54.67±4.509)%(P=0.025)和MMP-9表達(dá)升高增加(0.4767±0.07234)倍(P=0.038)。結(jié)論1.MiR-187在胃癌組織和人胃腺癌細(xì)胞SGC中表達(dá)明顯高于癌旁正常組織和人胃粘膜細(xì)胞GES,CD276在胃癌組織和人胃腺癌細(xì)胞SGC中表達(dá)明顯低于癌旁正常組織和人胃粘膜細(xì)胞GES。2.MiR-187和CD276的表達(dá)強(qiáng)度與胃癌患者淋巴結(jié)轉(zhuǎn)移個(gè)數(shù)、TNM分期、浸潤深度及Cerb B-2的表達(dá)相關(guān),而與患者的年齡、性別、腫瘤的部位、腫瘤大小及Ki67的表達(dá)無關(guān)。3.在胃癌組織和癌旁正常組織,miR-187和CD276之間存在線性負(fù)相關(guān),R2=0.84。4.抑制SGC細(xì)胞中miR-187的表達(dá)使胃癌細(xì)胞增殖受抑,侵襲和遷移能力下降。胃癌細(xì)胞中CD276表達(dá)升高,MMP-9表達(dá)下降。5.干擾SGC細(xì)胞中CD276的表達(dá)促進(jìn)胃癌細(xì)胞SGC增殖,胃癌細(xì)胞侵襲與遷移能力增高,MMP-9表達(dá)增高。
[Abstract]:Objective to investigate the effect of microRNA-187 (miR-187) expression in gastric carcinoma and gastric cancer proliferation, invasion and migration in vitro and its mechanism. Methods 24 in human gastric cancer tissues and adjacent normal tissues, the expression of miR-187 was detected in normal tissues of gastric cancer and adjacent, and explore its relationship with the clinicopathological parameters. The relationship between the expression of miR-187 detection the related gene CD276 in gastric cancer tissues and the analysis of both were inhibited the expression of miR-187 and CD276 human gastric cancer cell SGC-7901; miR-187 and CD276 expression of SGC-7901 cells on cell proliferation after change by CCK-8 method; the expression changes after SGC-7901 cell invasion and migration changes with miR-187 and CD276 Transwell assay; Western blot detection of SGC-7901 cells miR-187 and CD276 expression changes of matrix metalloproteinase -9 (matrix metalloproteinase-9, MMP- 9) expression changes. Results 1. real time fluorescence quantitative PCR results showed that the expression of miR-187 in gastric cancer tissue increased, and low expression in the adjacent (P0.001).Western blot showed that the decreased expression of CD276 in gastric cancer tissues, the expression of.MiR-187 and lymph node metastasis of gastric cancer patients and the number of high expression in adjacent tissues (P=0.046), TNM stage (P=0.015) and Cerb B-2 (P=0.013) expression, and the gender and age of patients, tumor size, tumor location, depth of invasion, independent expression and Ki67 (P 0.05).MiR-187 in lymph node metastasis of gastric cancer was 3, significantly stronger than the lymph node metastasis is less than 3 the expression of III and IV were Cerb and B-2 positive gastric cancer tissues, I and II and Cerb B-2 negative gastric cancer tissue.CD276 expression intensity and the number of lymph node metastasis (P=0.044), depth of invasion (P=0.042), TNM stage (P=0.001) and Cerb B-2 (P0.001) of the table 杈劇浉鍏，

本文編號：1604852