兰州大学机构库 >学院待认领
下调XIAP对胃癌细胞凋亡、增殖及细胞周期的影响
Alternative TitleThe effect of downregulating XIAP on apoptosis, proliferation and cell cycle
龙岩君
Thesis Advisor周永宁
2015-05-29
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
KeywordXIAP 胃癌 凋亡 细胞增殖 细胞周期
Abstract目的:前期研究发现miR-30a-5p在胃癌细胞株中低表达,并初步证实XIAP为miR-30a-5p的靶基因。本课题旨在探讨XIAP在人胃癌组织和胃癌细胞中的表达情况,以及下调XIAP后对胃癌细胞凋亡、增殖和细胞周期的影响及其可能的作用机制。 方法:收集54例胃癌患者的组织标本及其临床资料,用qRT-PCR法检测组织标本中XIAP mRNA的表达情况;以人正常胃黏膜上皮细胞GES-1为对照,采用qRT-PCR法和Western Blot检测胃癌细胞AGS、SGC-7901、MGC-803、BGC-823中XIAP mRNA和蛋白的表达情况。人工设计合成两对XIAP siRNA干扰序列瞬时转染以上5株细胞,以si-NC为对照,用同上方法检测XIAP mRNA和蛋白的表达水平及干扰效率。Annexin V/PI法、CCK8法及流式细胞术分别检测沉默XIAP后细胞凋亡、增殖及细胞周期的变化。 结果:XIAP在胃癌组织中的表达高于癌旁组织(P<0.001),且与免疫组化标记物Her-2的表达有相关性(P<0.01)。在胃癌细胞株AGS、SGC-7901、MGC-803、BGC-823中XIAP蛋白的表达均高于正常胃黏膜上皮细胞GES-1(P<0.05),但在AGS细胞中XIAP mRNA的相对表达量低于GES-1。瞬时转染以上5株细胞后, XIAP mRNA和蛋白的表达均较阴性对照序列组(si-NC)降低(P< 0.05/ P <0.01),干扰效率显著。下调XIAP后可增加胃癌细胞的凋亡率,呈时间依赖性抑制胃癌细胞的增殖(P < 0.05/ P <0.01),并诱导G0/G1和(或)G2/M期细胞阻滞。 结论: XIAP mRNA在胃癌组织中表达上调,且与免疫组化标记物Her-2的表达有相关性。XIAP蛋白在胃癌细胞株中表达上调,但与细胞内XIAP mRNA的表达水平有差异。下调XIAP后,可明显促进胃癌细胞凋亡,抑制细胞增殖,诱导细胞发生G0/G1和(或)G2/M期细胞阻滞。
Other AbstractObjective: Previous studies found that miR-30a-5p was lower expression in gastric cancer cell lines, and preliminary confirmed that XIAP was the target gene of miR-30a-5p.To investigate the expression of XIAP in human gastric cancer tissues and cells, and the effect of downregulating XIAP on apoptosis, proliferation and cell cycle in gastric cancer cells.Methods: 54 gastric cancer tissues compared to adjacent non-tumor tissues including their clinicopathological features were obtained,qRT-PCR was used to detect XIAP mRNA expression in tissue samples. The human gastric epithelial cell line GES-1 as a control, the expression of XIAP mRNA and protein in gastric cancer cell AGS, SGC-7901, MGC-803 and BGC-823 were detected by qRT-PCR and western blot. Two siRNA sequence were designed and synthesized artificially. After transient transfection the above cells, XIAP mRNA and protein were detected again to observe the expression level and interference efficiency compared with si-NC. Three groups of interference cells were stained with Annexin V/PI was used to analyze the effects on cells apoptosis by flow cytometry, proliferation viability was measured by CCK 8 assay, and cell cycle was tested by flow cytometry.Results: XIAP expression in gastric cancer tissues was significantly up-regulated compared to the adjacent non-tumor tissue (P<0.001), and associated with the expression of Her-2 tested by immunohistochemical stain(P<0.01). Cellular XIAP protein expression levels were increased in gastric cancer cell lines AGS, SGC-7901, MGC-803 and BGC-823, compared to gastric mucosa epithelial cells GES-1. However, the relative expression of XIAP mRNA in AGS cells was reduced. After transient transfection, expression level of XIAP mRNA and protein were significantly decreased than negative control group (si-NC) (P<0.05/P<0.01), presents the interference efficiency significantly. Downregulating XIAP aggrandizes apoptosis rates in gastric cancer cells, inhibits cell proliferation in a time-dependent manner (P<0.05/ P<0.01), and induced cell arrest in the G0/G1 and (or) G2/M stages.Conclusion: The up-regulated of XIAP was showed in gastric cancer tissues, and associated with the expression of Her-2 tested by immunohistochemical stain. XIAP protein expression were increased in gastric cancer cells, but were varied with the XIAP mRNA expression. Downregulating XIAP can promote apoptosis, inhibit cell proliferation, and induce G0/G1 and (or) G2/M cell arrest in gastric can...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222520
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
龙岩君. 下调XIAP对胃癌细胞凋亡、增殖及细胞周期的影响[D]. 兰州. 兰州大学,2015.
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