【作者】 杨静；

【导师】 贺军；

【作者基本信息】 南华大学 ， 临床医学（专业学位）， 2015， 硕士

【摘要】 目的:通过实时荧光定量PCR(q RT-PCR)技术从细胞和组织水平检测mi R-422a在不同肝细胞株及不同临床肝组织中的表达水平,分析其表达与肝细胞癌(HCC)患者临床病理特征的关系,为进一步深入研究mi R-422a在肝癌的发生、发展中的作用机制及意义奠定基础,同时为HCC生物学诊断、预后判断以及治疗提供新的手段依据,也为开发新的HCC药物提供基因靶点。方法:1.应用q RT-PCR技术检测mi R-422a基因在不同细胞株里的水平差异,细胞株分别是有侵袭能力的肝癌细胞和正常肝细胞;2.采用q RT-PCR技术检测临床标本中的30例HCC组织、30例相应癌旁组织及5例相对正常组织中mi R-422a的表达水平。3.分析mi R-422a的表达与HCC患者年龄、性别、肿瘤大小、肿瘤结节数目(单发或多发)、肿瘤分化程度(低、中、高分化)及静脉浸润等各项常见临床病理特征之间的关系。研究结果:q RT-PCR检测结果显示,mi R-422a在人正常肝细胞株(HL-7702)及三株不同侵袭转移能力的人HCC细胞株(侵袭力Hep G2<MHCC97-H<HCCLM3)中均有表达。mi R-422a在四种细胞中的表达水平从高到低依次为:HL-7702>Hep G2>MHCC97-H>HCCLM3,任意两株细胞株间表达水平的差异均有统计学意义(P<0.05)。1.q RT-PCR检测结果显示,mi R-422a在HCC组织、癌旁组织和正常肝组织中的表达存在差异,mi R-422a在HCC组织中表达低于相应癌旁组织及相对正常肝组织,各组织间表达水平两两比较均有统计学意义(P<0.05)。2.mi R-422a在肿瘤多发结节、低分化以及伴有静脉浸润的HCC组织中的表达分别明显高于单个结节(P=0.034)、高-中分化(P=0.033)和无静脉浸润(P=0.015)的HCC组织。mi R-422a其表达与性别、年龄、肿瘤直径、甲胎蛋白值、有无肝硬化及有无包膜无明显相关性(P>0.05)。结论:1.mi R-422a在人正常肝细胞株中表达上调,在HCC细胞株中较正常肝细胞株下调,且随着HCC细胞侵袭转移程度增加,其表达下调越明显。2.mi R-422a在HCC组织表达显著降低,且与HCC患者肿瘤多发结节、低分化程度以及伴有静脉浸润等临床病理特征密切相关。更多还原

【Abstract】 Background and Purpose of Study Hepatic cancer is one of the common Gastrointestinal Malignant Tumor in the world. However, hepatocellular carcinoma(HCC) constitutes the majority in hepatic cancer. The hepatic cancer patients have poor integral prognosis, especially for mal patients. In the countries where the patients having hepatic cancer increase for every year in the world, China is ranked in the first place, accounting for a half of overall incidence in the world. Since the early clinical symptoms of heptatic cancer are not typical. Most of the patients have entered in advanced hepatic cancer when they were diagnosed as hepatic cancer. For this reason, they have lost the opportunities of radical treatment including hepatectomy and liver transplantation. As for the hepatic cancer patients having the opportunities of surgical radical treatment, the five-year reoccurrence rate is 50% ~ 70% after hepatectomy. The main factors which cause lowering of survival rate and life quality are postoperative metastasis and reoccurrence of hepatic cancer. Hence so as a result, in order to increase the survival rate and improve life quality, we must improve diagnostic accuracy of hepatic cancer as far as possible on the onehand and adopt anti-relapse therapy against metastasis on the other hand. mi RNA(micro RNA) is micro-molecule RNA with wide distribution and non-coding and single chain. Its length is 18-25 nt. The studies have shown that the interaction between mi RNA cancer gene and cancer suppressor gene is the one of mechanisms of malignant tumor included in HCC. mi R-422 a acted as the member of mi RNA, the study of mi R-422 a is less relatively in the related studies of tumor. The results of the study have shown that mi R-422 a has played the important roles in occurrence process of laryngeal carcinoma, mammary cancer, colorectal adeno carcinoma, intestinal-type gastric carcinoma and osteosarcoma, and the apoptosis mechanisms after craniocerebral injury(such as Ischemic-reperfusion Injury, contusion of brain). In this experiment, it is intended to apply the real-time quantitative technology to determine the expression of mi R-422 a in hepatoma carcinoma cell and hepatocellular carcinoma tissue and analyze its expression and the relationships to clinic pathological characteristics with HCC patients ages, gender, tumor size, knot quantity of tumor, tumor differentiation and venous invasion, further study deeply that mi R-422 a plays the action mechanism in occurrence and development of hepatic cancer and lay the foundation of mean so as to provide the basis for mi RNA to play the new action in hepatic cancer and meanwhile to provide the basis for biological diagnosis, prognostic judgment and treatment and also to provide the gene target fordeveloping new medicines for treatment of hepatic cancer.Methods 1.By means of q RT-PCR Technology, to determine the expression differentials of mi R-422 a in human normal hepatocytes(HL-7702) and HCC Cell Strain with different invasion ability(Hep G2<MHCC97-H<JCCLM3) 2.By means of q RT-PCR technology, to determine the expression and correlation analysis of mi R-442 a in HCC tissue and corresponding adjacent tissues in 30 cases of HCC patients and in 5 cases of normal hepatic tissue 3.To analyze the expression of mi R-422 a and corelation between HCC clinic pathological characteristicsResults 1.The determination of q RT-PCR has shown that mi R-422 a has the expression in human normal hepatocytes(HL-7702) and HCC Cell Strain with the different invasion ability(Hep G2, MHCC97-H and HCCLM3). In comparison with optional two kinds cell stains(p<0.05), the differentials have the statistical significance. The expression of mi R-422 a in four kinds of cells: HL-7702>Hep G2>MHCC97-H>HCCLM3 2.The results of q RT-PCR has shown that the expression differentials of mi R-422 a in HCC tissue, corresponding adjacent tissues and normal tissue have the statistical significance(p<0.05) and comparing with comparisonhepatic tissue, mi R-422 a has the obvious downward in HCC tissue. 3.The expression of mi R-422 a in multiple nodules of tumor, poor differentiation and HCC tissue with venous invasion is distinctly higher than HCC tissue of the single nodule(p = 0.036), high-middle differentiation(p=0.035), non venous invasion(p = 0.0016). The expression of mi R-422 a has not obvious correlation with gender, age, diameter of tumor, AFP value, liver cirrhosis and peplos.Conclusion 1.mi R-422 a expresses upward in human normal hepatocytes, expresses downward in HCC cell strain and expresses obvious downward as HCC cell invasion and transfer increases. 2.The expression of mi R-422 a apparently decreases in HCC tissue and has the relationships to clinic pathological characteristics of HCC patients in multiple nodules of tumor, poor differentiation and accompanying venous invasion. Prompting that it may become a new biological marker of early diagnosis and prognosis assessment更多还原