本文選題：胃癌 + HEATR-1��； 參考：《皖南醫(yī)學院》2017年碩士論文

【摘要】：目的:運用兩株胃癌細胞BGC-823和AGS檢測HEATR-1(HEATR repeat containing-1)基因的表達,當其在胃癌細胞中高表達后,在胃癌組織和正常胃組織中檢測HEATR-1的表達情況;免疫組織化學方法觀察正常胃黏膜和胃癌組織石蠟切片中HEATR-1蛋白的表達。方法:根據(jù)在基因庫中檢索HEATR-1的m RNA核苷酸序列,設計、合成引物序列,胃癌細胞株經(jīng)培養(yǎng)后接種于六孔培養(yǎng)板中,行Q-PCR檢測胃癌細胞株BGC-823和AGS內(nèi)源性目的基因表達,確認其在胃癌細胞中高表達后,在術前明確診斷并手術的胃癌患者的胃癌組織行Q-PCR檢測HEATR-1的m RNA表達水平與正常組織之間表達,并對胃癌術后的石蠟切片行免疫組化染色,評估HEATR-1蛋白在石蠟切片的表達,并運用免疫組化評分方法,探討HEATR-1陽性表達與胃癌組織學類型的關系。結果:胃癌細胞株內(nèi)源性基因檢測,HEATR1基因高表達于兩株胃癌細胞AGS和BGC-823(AGSΔCt均值=7.10,BGC-823ΔCt均值=6.59,m RNA相對表達量分別為7.10±0.111、6.59±0.123);行20例胃癌患者的胃癌組織和陰性切緣行Q-PCR檢測HEATR-1的m RNA表達量,其中12例患者高表達,8例低表達,兩組間m RNA相對含量行配對t檢驗,P0.05,差異有統(tǒng)計學意義;選取胃癌和正常胃黏膜石蠟切片各12例,胃癌組織石蠟切片HEATR1蛋白明顯高表達于正常胃組織,并高表達于細胞膜和細胞質(zhì),用免疫組化評分方法,比較正常組和胃癌組之間免疫組化染色的差異,胃癌組評分(4.84±1.642),對照組評分(1.92±0.900),經(jīng)非配對t檢驗,P=0.032,P0.05,兩組間差異有統(tǒng)計學意義,以上結果,表明HEATR1高表達可能與胃癌的發(fā)生發(fā)展有關。結論:HEATR-1基因在胃癌細胞株和胃癌組織中高表達,其可能是胃癌發(fā)生的一個致病基因,并有望成為后期胃癌分子免疫治療的靶點。
[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 鹵0.111 ~ 6.59 鹵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 鹵1.642 and 1.92 鹵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.
【學位授予單位】：皖南醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.2

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本文編號：1982016