【作者】 陈琳；

【导师】 唐三元；

【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:通过检测DCLK1与mi R-217在胃癌组织和配对癌旁组织的表达;分析DCLK1与mi R-217的表达水平与胃癌临床病理参数间的关系,分析DCLK1与mi R-217在胃癌中表达的相关关系。探讨DCLK1与mi R-217的表达水平在胃癌中的意义。方法:应用生物信息学方法预测DCLK1的靶mi RNAs,并分析与其结合最稳定、特异性最好的靶mi RNA;收集49例新鲜的胃癌及配对的癌旁组织标本,一部分制作成石蜡切片,应用免疫组化技术检测DCLK1在胃癌组织与配对癌旁组织的表达情况;一部分用于提取总RNA,应用q RT-PCR检测DCLK1的靶mi RNA mi R-217在49例胃癌组织与配对癌旁组织的表达情况;分析DCLK1与mi R-217的表达水平与临床病理参数间的关系。分析DCLK1与mi R-217表达水平的相关性。结果:1.通过生物信息学方法预测发现了11个能与DCLK1结合的靶mi RNAs,并且通过进一步分析发现在这些靶mi RNAs中mi R-217与DCLK1结合的稳定性最高并且特异性也最好。2.应用免疫组化检测DCLK1在胃癌组织及其配对的癌旁组织的表达。结果显示DCLK1在胃癌组织中高表达率为73.5%(36/49),在配对的癌旁组织中的高表达率为38.8%(18/49),DCLK1在胃癌组织中高表达,两者具有显著统计学差异(P<0.05)。3.应用实时定量PCR检测胃癌组织和配对的癌旁组织mi R-217的表达。结果显示:与配对癌旁组织相比,mi R-217在胃癌组织中呈低表达的有77.6%(38/49),呈高表达的有22.4%(11/49),mi R-217在胃癌组织中的表达是下调的;mi R-217在胃癌组织中的相对表达量为0.17180±3.346363,在配对癌旁组织中相对表达量为1.32878±2.371940。mi R-217在胃癌组织中的表达水平低于配对癌旁组织,改变具有显著统计学差异(P<0.05)。4.DCLK1的表达水平与胃癌的分化程度、淋巴结转移、TNM分期显著相关(P<0.05),而与性别、年龄、肿瘤直径、肿瘤浸润的深度无关(P>0.05)。5.mi R-217的表达水平与胃癌的分化程度、淋巴结转移、TNM分期显著相关(P<0.05),而与性别、年龄、肿瘤直径、肿瘤浸润的深度无关(P>0.05)。6.Spearman秩检验相关分析显示在胃癌组织中DCLK1与mi R-217的表达之间呈负相关(r=-0.675,P<0.01)。结论:1.DCLK1的高表达和mi R-217低表达可能与胃癌的发生发展有关。2.DCLK1与mi R-217在胃癌中的表达呈负相关,DCLK1与mi R-217在胃癌中的表达水平有望成为评估判断胃癌恶性程度的标志物。更多还原

【Abstract】 Objective:The expression of DCLK1 and mi R-217 was detected in gastric cancer tissues and adjacent tissues. Additionally the relationship was analyzed between clinicopathological parameters and the expression level of DCLK1 and mi R-217 in gastric cancer tissues. The correlation of DCLK1 and mi R-217 was also analyzed at the same time. We discussed the clinical Significance that is related to the expression level of DCLK1 and mi R-217 in the gastric cancer. Methods:Target mi RNAs of DCLK1 were predicted by bioinformatics methods, and analyzed to find the mi RNA which targets DCLK1 with the best stability and specificity.The tissues of gastric cancer and adjacent gastric were obtained from 49 patients. The samples were divided into two fractions,one was made into paraffin in order to research the level of DCLK1. The expression of DCLK1 were detected by immunohistochemistry in gastric cancer tissues and adjacent tissues. another was extracted total RNA from the tissues of gastric cancer and adjacent tissues. The expression of mi R-217 in 49 tissues of gastric cancer and adjacent tissues was detected by Quantificational real-time polymerase chain reaction. The relationships was analyzed by statistical analysis between expression level of DCLK1 and mi R-217 and clinicalpathological parameters of gastric cancer.The correlation of DCLK1 and mi R-217 was analyzed. Results:1、11 target mi RNAs of DCLK 1 which could bind to DCLK1 were obtained by bioinformatics methods. The analyzed result indicated that mi R-217 is the one which targets DCLK1 with the best stability and specificity by Cross analysis.2、The expression of DCLK1 was detected by immunohistochemistry in gastric cancer tissues and adjacent tissues. The result show that the high expression ratio of DCLK1 in gastric cancer tissues was 73.5%(36/49), the high expression ratio of DCLK1 in adjacent tissues was 38.8%(18/49). The expression of DCLK1 was high in gastric cancer tissues, the difference was stastically significant(P<0. 05).3、The expression of mi R-217 was detected by Quantificational real-time polymerase chain reaction. compared with adjacent tissues,The result showed that the low expression ratio of mi R-217 in gastric cancer tissues was 77.6%(38/49), high expression ratio of mi R-217 was 22.4%(11/49). The expression of mi R-217 was decreased in gastric cancer tissues. The relative intensity of mi R-217 in gastric cancer tissues was( 0.17180±3.346363), in adjacent tissues was(1.32878±2.371940). The expression of mi R-217 was significantly lower in gastric cancer tissues than that in adjacent tissues, the difference was stastically significant(P<0.05).4、The expression level of DCLK1 in gastric carcinoma was associated with differentiation, lymph node metastases TNM stages(P<0.05),but not with gender,age, tumor size and depth of invasion(P>0.05).5、The expression level of mi R-217 in gastric cancer was associated with differentiation, lymph node metastases,TNM stages(P<0.05),but not with gender, age, tumor size and depth of invasion(P>0.05).6、Spearman analysis indicated that there was a significantly ne gative correlation between DCLK1 and mi R-217.(r=-0.675,P<0.01). Conclusions:1、The high expression of DCLK1 and low expression of mi R-217 were associated with the occurrence and development of gastric cancer.2、The expression of DCLK1 and mi R-217 was negative correlation.The expression of DCLK1 and mi R-217 in gastric cancer is expected to become markers that can assess degree of malignancy.更多还原