HEATR-1在胃癌中的表达及其相关性研究

发布时间：2018-06-05 12:38

本文选题：胃癌 + HEATR-1　；参考：《皖南医学院》2017年硕士论文

【摘要】：目的:运用两株胃癌细胞BGC-823和AGS检测HEATR-1(HEATR repeat containing-1)基因的表达,当其在胃癌细胞中高表达后,在胃癌组织和正常胃组织中检测HEATR-1的表达情况;免疫组织化学方法观察正常胃黏膜和胃癌组织石蜡切片中HEATR-1蛋白的表达。方法:根据在基因库中检索HEATR-1的m RNA核苷酸序列,设计、合成引物序列,胃癌细胞株经培养后接种于六孔培养板中,行Q-PCR检测胃癌细胞株BGC-823和AGS内源性目的基因表达,确认其在胃癌细胞中高表达后,在术前明确诊断并手术的胃癌患者的胃癌组织行Q-PCR检测HEATR-1的m RNA表达水平与正常组织之间表达,并对胃癌术后的石蜡切片行免疫组化染色,评估HEATR-1蛋白在石蜡切片的表达,并运用免疫组化评分方法,探讨HEATR-1阳性表达与胃癌组织学类型的关系。结果:胃癌细胞株内源性基因检测,HEATR1基因高表达于两株胃癌细胞AGS和BGC-823(AGSΔCt均值=7.10,BGC-823ΔCt均值=6.59,m RNA相对表达量分别为7.10±0.111、6.59±0.123);行20例胃癌患者的胃癌组织和阴性切缘行Q-PCR检测HEATR-1的m RNA表达量,其中12例患者高表达,8例低表达,两组间m RNA相对含量行配对t检验,P0.05,差异有统计学意义;选取胃癌和正常胃黏膜石蜡切片各12例,胃癌组织石蜡切片HEATR1蛋白明显高表达于正常胃组织,并高表达于细胞膜和细胞质,用免疫组化评分方法,比较正常组和胃癌组之间免疫组化染色的差异,胃癌组评分(4.84±1.642),对照组评分(1.92±0.900),经非配对t检验,P=0.032,P0.05,两组间差异有统计学意义,以上结果,表明HEATR1高表达可能与胃癌的发生发展有关。结论:HEATR-1基因在胃癌细胞株和胃癌组织中高表达,其可能是胃癌发生的一个致病基因,并有望成为后期胃癌分子免疫治疗的靶点。
[Abstract]:Objective: to detect the expression of HEATR-1(HEATR repeat containing-1 (HEATR-1(HEATR repeat containing-1) gene in two gastric cancer cell lines (BGC-823 and AGS), and to detect the expression of HEATR-1 in gastric cancer tissues and normal gastric tissues after high expression of HEATR-1(HEATR repeat containing-1 gene. The expression of HEATR-1 protein in paraffin sections of normal gastric mucosa and gastric carcinoma was observed by immunohistochemical method. Methods: according to the m RNA nucleotide sequence of HEATR-1 in gene bank, the primer sequence was designed and synthesized, and the gastric cancer cell line was cultured and inoculated into a six-well culture plate. The expression of endogenous target gene of BGC-823 and AGS in gastric cancer cell line was detected by Q-PCR. To confirm its high expression in gastric cancer cells, Q-PCR was used to detect the expression of m RNA in gastric cancer tissues of patients with gastric cancer diagnosed and operated on before operation, and the expression of m RNA in normal tissues was detected by Q-PCR, and the paraffin sections of gastric cancer were stained with immunohistochemistry. To evaluate the expression of HEATR-1 protein in paraffin sections and to explore the relationship between the positive expression of HEATR-1 and the histological types of gastric cancer by immunohistochemistry. Results: the expression of pHEATR1 gene was high in AGS and BGC-823(AGS 螖 Ct cells of two gastric cancer cell lines, and the relative expression of HEATR-1 was 7.10 卤0.111 ~ 6.59 卤0.123 m RNA in the mean value of BGC-823 螖 Ct, respectively, and Q-PCR was used to detect the expression of HEATR-1 in 20 gastric cancer tissues and negative margin of gastric cancer. There were 12 cases of high expression and 8 cases of low expression, the relative content of m RNA in the two groups was matched t test (P0.05), the difference was statistically significant, 12 cases of gastric cancer and 12 cases of normal gastric mucosa paraffin sections were selected. HEATR1 protein in paraffin sections of gastric cancer tissues was significantly overexpressed in normal gastric tissues, and also in cell membrane and cytoplasm. Immunohistochemistry was used to compare the difference of immunohistochemical staining between normal and gastric cancer groups. The scores of gastric cancer group and control group were 4.84 卤1.642 and 1.92 卤0.900, respectively. The difference between the two groups was statistically significant. The above results indicated that the high expression of HEATR1 might be related to the occurrence and development of gastric cancer. Conclusion the expression of the gene is highly expressed in gastric cancer cell lines and gastric cancer tissues. It may be a pathogenic gene of gastric cancer and may be a target of molecular immunotherapy for late gastric cancer.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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