MicroRNA-187在胃癌中的表达及对胃癌细胞增殖和侵袭影响的研究

发布时间：2018-03-13 04:34

  本文选题：microRNA-187　切入点：CD276　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的探讨microRNA-187(miR-187)在胃癌中的表达及对胃癌增殖、侵袭和迁移的影响及其机制。方法收集24对人胃癌组织和癌旁正常组织,检测miR-187在胃癌组织和癌旁正常组织的表达,探讨其与各临床病理指标之间关系。检测miR-187相关基因CD276在胃癌组织组织中的表达并分析两者的相关性。分别抑制人胃腺癌细胞SGC-7901中miR-187和CD276的表达;用CCK-8法检测SGC-7901细胞中miR-187和CD276表达发生改变后对细胞增殖的影响;用Transwell法检测miR-187和CD276表达发生改变后SGC-7901细胞侵袭性及迁移性的改变;用Western blot检测SGC-7901细胞miR-187和CD276表达发生改变后基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达改变。结果1.实时荧光定量PCR结果显示miR-187在胃癌组织中表达升高,而在癌旁组织中低表达(P0.001)。Western blot检测证实CD276在胃癌组织中表达降低,而在癌旁组织中高表达。miR-187的表达强度与胃癌患者淋巴结转移个数(P=0.046)、TNM分期(P=0.015)及Cerb B-2(P=0.013)的表达相关,而与患者的年龄、性别、肿瘤的部位、肿瘤大小、浸润深度及Ki67的表达无关(P均0.05)。miR-187在胃癌淋巴结转移≥3枚、III期和IV期及Cerb B-2阳性的胃癌组织中表达明显强于淋巴结转移小于3枚、I期和II期及Cerb B-2阴性的胃癌组织。CD276的表达强度与淋巴结转移个数(P=0.044)、浸润深度(P=0.042)、TNM分期(P=0.001)及Cerb B-2(P0.001)的表达相关,而与患者的年龄、性别、肿瘤的部位、肿瘤大小及Ki67的表达无关(P均0.05)。CD276在肿瘤淋巴结转移≥3枚、T3~T4、III期和IV期及Cerb B-2阳性的胃癌组织中表达明显低于淋巴结转移小于3枚、T1~T2、I期和II期及Cerb B-2阴性的胃癌组织。2.与癌旁正常组织比较,miR-187在I期、II期、III期和IV期胃癌组织表达进行性升高,分别是正常组织的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期胃癌组织表达降低,分别是正常组织的0.68倍、0.71倍、0.67倍和0.61倍(P0.01);miR-187在胃腺癌细胞SGC中的表达是胃癌粘膜上皮细胞GES的5.5倍(P0.001)。3.胃癌组织中miR-187表达与CD276表达相关系数R2值为0.840,表示在此胃癌标本中miR-187表达量可以解释CD276表达量变异性的76.63%,另外23.37%的变异不能用miR-187表达量解释。4.与对照组(miR-187 inhibitor-control组)比较,miR-187表达抑制组(miR-187inhibitor)SGC增殖活性较对照组降低(60.67±11.02)%(P=0.003)、侵袭性较对照组降低(60.67±11.02)%(P=0.001)和迁移能力较对照组降低(66.33±10.6)%(P0.001);miR-187 inhibitor组CD276表达增加(1.047±0.06506)倍(P0.001)和MMP-9表达降低(0.2667±0.06028)倍(P0.001)。5.与对照组(CD276-siRNA-control组)比较,CD276表达干扰组(CD276-siRNA)SGC增殖活性为对照组的(1.823±0.2577)倍(P=0.002)、侵袭性为对照组的(1.843±0.3296)倍(P0.001)与迁移性为对照组的(1.66±0.1453)倍(P0.001);CD276-siRNA组CD276表达降低(54.67±4.509)%(P=0.025)和MMP-9表达升高增加(0.4767±0.07234)倍(P=0.038)。结论1.MiR-187在胃癌组织和人胃腺癌细胞SGC中表达明显高于癌旁正常组织和人胃粘膜细胞GES,CD276在胃癌组织和人胃腺癌细胞SGC中表达明显低于癌旁正常组织和人胃粘膜细胞GES。2.MiR-187和CD276的表达强度与胃癌患者淋巴结转移个数、TNM分期、浸润深度及Cerb B-2的表达相关,而与患者的年龄、性别、肿瘤的部位、肿瘤大小及Ki67的表达无关。3.在胃癌组织和癌旁正常组织,miR-187和CD276之间存在线性负相关,R2=0.84。4.抑制SGC细胞中miR-187的表达使胃癌细胞增殖受抑,侵袭和迁移能力下降。胃癌细胞中CD276表达升高,MMP-9表达下降。5.干扰SGC细胞中CD276的表达促进胃癌细胞SGC增殖,胃癌细胞侵袭与迁移能力增高,MMP-9表达增高。
[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 杈剧浉鍏，

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