發(fā)布時(shí)間：2018-06-05 12:38

  本文選題：胃癌 + HEATR-1�。� 參考：《皖南醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:運(yùn)用兩株胃癌細(xì)胞BGC-823和AGS檢測(cè)HEATR-1(HEATR repeat containing-1)基因的表達(dá),當(dāng)其在胃癌細(xì)胞中高表達(dá)后,在胃癌組織和正常胃組織中檢測(cè)HEATR-1的表達(dá)情況;免疫組織化學(xué)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(dá)。方法:根據(jù)在基因庫(kù)中檢索HEATR-1的m RNA核苷酸序列,設(shè)計(jì)、合成引物序列,胃癌細(xì)胞株經(jīng)培養(yǎng)后接種于六孔培養(yǎng)板中,行Q-PCR檢測(cè)胃癌細(xì)胞株BGC-823和AGS內(nèi)源性目的基因表達(dá),確認(rèn)其在胃癌細(xì)胞中高表達(dá)后,在術(shù)前明確診斷并手術(shù)的胃癌患者的胃癌組織行Q-PCR檢測(cè)HEATR-1的m RNA表達(dá)水平與正常組織之間表達(dá),并對(duì)胃癌術(shù)后的石蠟切片行免疫組化染色,評(píng)估HEATR-1蛋白在石蠟切片的表達(dá),并運(yùn)用免疫組化評(píng)分方法,探討HEATR-1陽(yáng)性表達(dá)與胃癌組織學(xué)類型的關(guān)系。結(jié)果:胃癌細(xì)胞株內(nèi)源性基因檢測(cè),HEATR1基因高表達(dá)于兩株胃癌細(xì)胞AGS和BGC-823(AGSΔCt均值=7.10,BGC-823ΔCt均值=6.59,m RNA相對(duì)表達(dá)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(cè)HEATR-1的m RNA表達(dá)量,其中12例患者高表達(dá),8例低表達(dá),兩組間m RNA相對(duì)含量行配對(duì)t檢驗(yàn),P0.05,差異有統(tǒng)計(jì)學(xué)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(dá)于正常胃組織,并高表達(dá)于細(xì)胞膜和細(xì)胞質(zhì),用免疫組化評(píng)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(píng)分(4.84±1.642),對(duì)照組評(píng)分(1.92±0.900),經(jīng)非配對(duì)t檢驗(yàn),P=0.032,P0.05,兩組間差異有統(tǒng)計(jì)學(xué)意義,以上結(jié)果,表明HEATR1高表達(dá)可能與胃癌的發(fā)生發(fā)展有關(guān)。結(jié)論:HEATR-1基因在胃癌細(xì)胞株和胃癌組織中高表達(dá),其可能是胃癌發(fā)生的一個(gè)致病基因,并有望成為后期胃癌分子免疫治療的靶點(diǎn)。
[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(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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