甲苯磺酸索拉非尼治疗晚期原发性肝细胞癌患者的临床病理特征、生存预后分析及低氧诱导其耐药机制研究

发布时间：2018-04-20 15:21

本文选题：肝细胞癌(HCC) + 甲苯磺酸索拉非尼　；参考：《中国人民解放军医学院》2016年博士论文

【摘要】：目的：回顾性收集并研究于我院口服甲苯磺酸索拉非尼片的晚期原发性肝细胞肝癌患者的临床及病理资料,统计分析与甲苯磺酸索拉非尼片临床疗效相关的预后因素：探索低氧环境诱发的甲苯磺酸索拉非尼耐药机制。材料与方法：回顾性收集了2005年1月至2013年1月期间于我院口服甲苯磺酸索拉非尼片的晚期HCC患者的临床病理资料,收集这些患者的肝癌组织蜡块标本并切片,应用SPSS19.9统计软件对生存预后进行单因素分析及多因素分析;培养肝癌细胞系,进行Western Blot、PCI、CCK8毒性检测、HIF1SiRNA转染、免疫荧光染色等。结果：(1)74例患者口服甲苯磺酸索拉非尼,中位随访时间为535天；其中64例为男性患者,10例为女性患者(6.4：1),中位年龄62岁,丙型肝炎既往史、较高的KPS评分、无肿瘤家族史的患者中位PFS相对较长;丙型肝炎既往史、AFP400, KPS评分≥90,高分化癌,不伴有吸烟史、饮酒史及肿瘤家族史的患者中位生存期较长;肿瘤家族史、乙型肝炎、KPS评分70-80、低分化癌是生存的不良预后因素;饮酒既往史、乙型肝炎、KPS评分70-80是与甲苯磺酸索拉非尼片疗效相关的不良预后因素；(2)口服甲苯磺酸索拉非尼片疗效较差的临床晚期HCC患者,免疫荧光染色显示组织标本中HIF 1α及YAP在细胞核内聚集,胞核阳性率较高,而临床获益明显的患者肝癌组织中,HIF 1α及YAP主要集中在细胞质;(3)在正常培养的肝癌细胞中,HIF1α及YAP均有少量表达,但重合率较低且主要集中在细胞质,细胞核阳性率低;(4)肝癌细胞在常氧环境下加用甲苯磺酸索拉非尼,HIF 1α及YAP主要集中在细胞质,细胞核阳性率进一步降低;(5)肝癌细胞在低氧环境下加用甲苯磺酸索拉非尼,HIF 1α及YAP共同向核内迁移,细胞核阳性率升高;(6)低氧环境下,YAP、HIF、1α表达升高且主要集中在细胞核,提示低氧环境可能促成肝癌细胞对甲苯磺酸索拉非尼产生抵抗力,促使产生耐药；(7)当YAP主要集中在肝癌细胞核时,EMT相关指标E-钙粘附减少,N-钙粘附素、波形蛋白表达水平相应增多,预示着YAP向细胞核内迁移现象与肝癌细胞的恶性程度及侵袭性密切相关;(8)正常培养的肝癌细胞中,细胞毒性检测提示低氧状态可能抵抗甲苯磺酸索拉非尼抗肿瘤效应,逆向促进肿瘤细胞增殖生长;(9) HIF1SiRNA转染肝癌细胞,当HIF la表达下调后,对其进行低氧及甲苯磺酸索拉非尼的处理,YAP的表达水平及向细胞核的迁移现象同前无明显变化,但是细胞毒性检测实验显示,肝癌细胞对索拉非尼的耐受性有所下降；(10) HIF 1α及YAP蛋白在低氧诱导的甲苯磺酸索拉非尼耐药机制中起到了一定的协同作用。结论：(1)通过对口服甲苯磺酸索拉非尼片的晚期HCC患者的临床病理资料及生存预后分析,我们发现肝炎病史、KPS评分、肿瘤家族史等临床因素与患者的生存及甲苯磺酸索拉非尼片的疗效存在密切关系,对该类临床患者的治疗及预后具有一定的指导意义;(2)低氧环境能够抵抗甲苯磺酸索拉非尼部分抗肿瘤效应,诱导其耐药的产生;(3)低氧环境下,YAP向细胞核内迁移迁移现象与耐药及预后有着重要关系;(4)HIF1SiRNA转染的肝癌细胞对索拉菲尼的敏感性有所增加；(5) HIF la及YAP在低氧诱导的甲苯磺酸索拉非尼耐药机制中可能起到了一定的协同作用。
[Abstract]:Objective: To retrospectively collect and study the clinical and pathological data of patients with advanced primary hepatocellular carcinoma (HCC) in our hospital, and to analyze the prognostic factors associated with the clinical efficacy of Sorafenib Tosylate Tablets: explore the mechanism of ssorafeni resistance induced by the hypoxic environment. Materials and methods: The clinicopathological data of late HCC patients in our hospital from January 2005 to January 2013 were collected, and the paraffin specimens of these patients were collected and sliced. A single factor analysis and multi factor analysis were used to analyze the survival prognosis with SPSS19.9 statistical software, and to develop the hepatoma cell line for West. Ern Blot, PCI, CCK8 toxicity test, HIF1SiRNA transfection, immunofluorescence staining and so on. Results: (1) 74 patients were taken orally with sorafenib toluene sulfonic acid, the median follow-up time was 535 days, of which 64 were male patients, 10 were female patients (6.4:1), middle age 62 years, hepatitis C history, higher KPS score, and no family history of tumor. The position of PFS was relatively long; the past history of hepatitis C, AFP400, KPS score was more than 90, high differentiated cancer, no history of smoking, drinking history and family history of cancer had a longer median survival period; the family history of cancer, hepatitis B, KPS score, and low differentiated carcinoma were the poor prognostic factors of survival; drinking past history, hepatitis B, and KPS score 70-80 were with toluene sulphur. The adverse prognostic factors associated with acid sorafenib (2) the late clinical HCC patients with poor therapeutic efficacy of oral Sorafenib Tosylate Tablets showed that HIF 1 alpha and YAP were aggregated in the nucleus and the positive rate of the nucleus was higher in the tissue specimens. The HIF 1 alpha and YAP were mainly concentrated in the liver cancer tissues of the patients with significant clinical benefits. Cytoplasm; (3) in normal cultured hepatoma cells, HIF1 A and YAP had a small amount of expression, but the recoincidence rate was low and mainly concentrated in the cytoplasm, and the positive rate of the nucleus was low; (4) the hepatoma cells were added with toluene sulfonic acid Sola Fini, HIF 1 A and YAP mainly in the cytoplasm, and the positive rate of the nucleus was further reduced; (5) hepatoma cells In the hypoxic environment, Sola Fini, HIF 1 alpha and YAP were migrated to the nucleus, and the positive rate of the nucleus increased; (6) the expression of YAP, HIF, 1 alpha was increased and mainly concentrated in the nucleus under the environment of hypoxia, suggesting that the hypoxia environment may contribute to the resistance of the hepatoma cells to toluene sulfonic acid and induce resistance; (7) when YAP main. To concentrate on the cell nucleus of liver cancer, the EMT related index E- calcium adherence decreases, N- calcin and vimentin expression increase correspondingly, indicating that the migration of YAP into the nucleus is closely related to the malignancy and invasiveness of the hepatoma cells. (8) the cytotoxicity test suggests that hypoxia may resist toluene in normal cultured hepatoma cells. The effect of sorafeni sulfonate on tumor cell proliferation and growth of tumor cells; (9) HIF1SiRNA transfected hepatoma cells, when the expression of HIF La was downregulated, it was treated with hypoxia and sorafenib toluene sulfonic acid. The expression level of YAP and the migration of the nucleus to the nucleus had no obvious change, but the cytotoxicity test showed that the liver cancer was fine. The tolerance to sorafenib decreased; (10) HIF 1 alpha and YAP protein had a synergistic effect in the mechanism of low oxygen induced sorafenib resistance in toluene sulfonic acid. Conclusion: (1) we found the history of hepatitis, KP, through the analysis of the clinicopathological materials and survival prognosis of the advanced HCC patients with oral Sorafenib Tosylate Tablets. S score, the family history of tumor and other clinical factors are closely related to the survival of the patients and the curative effect of Sorafenib Tosylate Tablets, and have certain guiding significance for the treatment and prognosis of this kind of clinical patients. (2) the hypoxia environment can resist the Sola Fini partial swelling tumor effect of toluene sulfonic acid and induce the production of its resistance; (3) Y in the hypoxia environment. The migration and migration of AP into the nucleus has an important relationship with drug resistance and prognosis. (4) the sensitivity of HIF1SiRNA transfected hepatoma cells to Sola Feeney is increased; (5) HIF La and YAP may play a certain synergistic effect in the mechanism of low oxygen induced sorafenib toluene sulfonic acid.

【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7

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