本文選題：ALCAM + 肝細胞癌�。� 參考：《重慶醫(yī)科大學》2016年碩士論文

【摘要】：背景活化白細胞黏附分子(Activated Leukocyte Cell Adhesion Molecule,ALCAM)是免疫球蛋白基因超家族成員,主要表達在白細胞和胸腺上皮細胞。ALCAM為I型跨膜蛋白,其通過異嗜性黏附形式(ALCAM-CD6)和同嗜性黏附形式(ALCAM-ALCAM)介導細胞間黏附。既往研究表明ALCAM與腫瘤侵襲與轉(zhuǎn)移密切相關(guān),參與多種腫瘤的發(fā)生發(fā)展。目的本研究探討ALCAM表達與肝細胞癌的相關(guān)性以及其在肝細胞癌發(fā)生發(fā)展中的作用。方法1.RT-PCR及免疫組化方法檢測ALCAM基因在52例肝細胞癌組織及相應(yīng)癌旁組織中的表達,分析ALCAM的表達與肝細胞癌臨床病理特征的相關(guān)性。2.利用p SES-HUS系統(tǒng)構(gòu)建特異性ALCAM基因的si RNA腺病毒載體p SES-ALCAM-si RNA,測序證實基因序列正確后包裝成腺病毒si RNA并轉(zhuǎn)染肝癌細胞株HepG2,應(yīng)用RT-PCR和Western blot檢測si RNA對肝癌細胞HepG2中ALCAM基因的干擾效果。3.MTT及流式細胞技術(shù)檢測干擾ALCAM表達后HepG2細胞增殖及凋亡情況;Transwell實驗檢測HepG2遷移、侵襲能力改變。結(jié)果1.RT-PCR結(jié)果顯示與癌旁正常組織相比較,ALCAM在肝細胞癌組織中的高表達(P0.01)。免疫組化與臨床資料統(tǒng)計分析進一步證實ALCAM與腫瘤分化程度(P=0.024),肝內(nèi)轉(zhuǎn)移(P=0.019)呈負相關(guān),肝癌組織中ALCAM異常低表達提示腫瘤分化程度低,易轉(zhuǎn)移;而與患者性別,年齡,術(shù)前AFP值,腫瘤直徑及臨床TNM分期無明顯相關(guān)性(P0.05)。2.構(gòu)建腺病毒p SES-ALCAM-si RNA并轉(zhuǎn)染HepG2,RT-PCR和Western blot顯示ALCAM在基因及蛋白表達水平均被明顯抑制。3.通過si RNA干擾肝癌細胞株HepG2中ALCAM基因表達,MTT及流式細胞術(shù)檢測表明干擾ALCAM表達后肝癌細胞株HepG2增殖受到抑制(P0.05),凋亡率增加(P0.05);而Transwell實驗發(fā)現(xiàn)ALCAM干擾后HepG2遷移、侵襲能力上升(P0.05)。結(jié)論1.ALCAM在肝細胞癌組織中的高表達,并且與肝癌分化程度,肝內(nèi)轉(zhuǎn)移有相關(guān)性。這些數(shù)據(jù)表明ALCAM可能作為一個新指標診斷肝癌并且提示腫瘤分化程度及轉(zhuǎn)移風險。2.腺病毒si RNA成功構(gòu)建并可特異性干擾HepG2肝癌細胞株ALCAM基因表達,為后繼實驗進一步揭示ALCAM與肝癌的關(guān)系做準備。3.干擾ALCAM基因可抑制肝癌細胞HepG2增殖,促進其凋亡;然而HepG2肝癌細胞在ALCAM基因表達下調(diào)后體外遷移和侵襲能力增強,這意味著ALCAM基因在肝癌發(fā)生發(fā)展中有著多方面的作用,有重要研究價值。
[Abstract]:The background activated leukocyte adhesion molecule (Activated Leukocyte Cell Adhesion Molecule, ALCAM) is a member of the immunoglobulin gene superfamily, mainly expressed in the leukocyte and thymic epithelial cells.ALCAM as a I type transmembrane protein, which mediates the intercellular adhesion through the heterophil adhesion form (ALCAM-CD6) and the isophil adhesion form (ALCAM-ALCAM). The study shows that ALCAM is closely related to tumor invasion and metastasis and participates in the occurrence and development of various tumors. Objective to investigate the relationship between ALCAM expression and hepatocellular carcinoma and its role in the development of hepatocellular carcinoma. Methods 1.RT-PCR and immunohistochemical methods were used to detect the ALCAM gene in 52 hepatocellular carcinoma tissues and corresponding Para cancerous tissues. The correlation between the expression of ALCAM and the clinicopathological features of hepatocellular carcinoma.2. using the P SES-HUS system to construct the Si RNA adenovirus vector p SES-ALCAM-si RNA of the specific ALCAM gene, and the sequence of DNA sequencing confirmed that the gene sequence was correctly packaged into adenovirus Si RNA and transfected to the liver cancer cell line. Interference effect on ALCAM gene in hepatoma cell HepG2.3.MTT and flow cytometry to detect the proliferation and apoptosis of HepG2 cells after interference of ALCAM expression; Transwell test was used to detect HepG2 migration and invasion ability. Results 1.RT-PCR results showed high expression of ALCAM in hepatocellular carcinoma tissues compared with normal tissues adjacent to cancer (P0.01). The statistical analysis of histochemical and clinical data further confirmed that ALCAM was negatively correlated with the degree of tumor differentiation (P=0.024) and intrahepatic metastasis (P=0.019). The abnormal low expression of ALCAM in the hepatocellular carcinoma showed that the tumor differentiation was low and easy to transfer, but there was no significant correlation between the sex, age, preoperative AFP, the diameter of the tumor and the clinical TNM staging (P0.05).2. construction gland. The virus P SES-ALCAM-si RNA was transfected with HepG2, RT-PCR and Western blot showed that the expression level of ALCAM in the gene and protein expression was obviously inhibited by Si RNA interference in the hepatocellular carcinoma cell line, and the proliferation of the hepatocellular carcinoma cell line was inhibited and the apoptosis rate increased after the flow cytometry. 05); and the Transwell experiment found that HepG2 migration and invasion ability increased after ALCAM interference (P0.05). Conclusion 1.ALCAM is highly expressed in hepatocellular carcinoma and is associated with the degree of differentiation and intrahepatic metastasis of HCC. These data suggest that ALCAM may be a new marker for the diagnosis of liver cancer and that the degree of differentiation and metastasis risk of.2. adenosis may be suggested. Si RNA was successfully constructed and specifically interfered with the expression of ALCAM gene in HepG2 cell line, which further revealed the relationship between ALCAM and HCC by preparing.3. interference ALCAM gene to inhibit the proliferation of hepatoma cell HepG2 and promote its apoptosis. However, the migration and invasion ability of HepG2 hepatoma cells was enhanced in vitro after the expression of ALCAM gene was down. This means that ALCAM gene plays an important role in the occurrence and development of liver cancer and has important research value.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R735.7

【參考文獻】

相關(guān)期刊論文 前3條

1 Sonia Pascual;Iván Herrera;Javier Irurzun;;New advances in hepatocellular carcinoma[J];World Journal of Hepatology;2016年09期

2 孫力超;李智峰;李闖;韓璐璐;楊治華;冉宇靚;;ALCAM在食管癌中的表達及其功能研究[J];中國腫瘤;2011年06期

3 ;Hepatocellular Carcinoma-Cause,Treatment and Metastasis[J];World Journal of Gastroenterology;2001年04期

，

本文編號：1914327