摘要
目的探讨微小RNA(miRNA,miR)-200a在前列腺癌组织中的表达及与临床参数的相关性,并在体内研究miR-200a对裸鼠移植瘤生长的影响及机制。方法应用荧光原位杂交(FISH)技术检测2018年1月至2019年8月在徐州医科大学附属医院接受手术切除的54例经病理证实的前列腺癌的组织标本及前列腺增生组织标本中miR-200a的表达,并分析与病理分级、Gleason评分、前列腺特异抗原(PSA)水平、年龄之间的相关性;建立小鼠皮下移植瘤模型(购自上海西普尔-必凯实验动物有限公司),分3组:转染miR-200a高表达质粒的DU145细胞组、转染表达miR-200a+特异性核基质结合区结合蛋白1(SATB1)的DU145细胞质粒组、未转染的DU145细胞组,取0.2 ml细胞悬液接种于小鼠右后臀部皮下,每隔5 d测量皮下移植瘤体积,绘制裸鼠移植瘤生长曲线,采用χ^2检验及t检验,分析3组移植瘤体积的统计学差异;免疫组织化学和蛋白质印迹法(Western blot)法检测移植瘤SATB1的表达变化。多样本间比较采用单因素方差分析,组间比较采用t检验。结果miR-200a在前列腺癌组织中的阳性率为27.8%(15/54),在前列腺增生组织中的阳性率为90.0%(9/10),差异有统计学意义(χ^2=11.409,P<0.05),且前列腺癌组织中miR-200a表达强度与前列腺癌的病理分级和临床分期及转移呈负相关,接种细胞后,对照组的移植瘤的SATB1的表达量高于miR-200a高表达组,转染miR-200a高表达质粒组的移植瘤最终体积为(615.39±31.05)mm^3,而未转染的DU145细胞组瘤体体积为(1489.09±192.54)mm^3,差异有统计学意义(t=7.759,P<0.05),说明在体内实验裸鼠皮下成瘤中miR-200a能够显著抑制前列腺癌裸鼠移植瘤的生长,且免疫组织化学实验结果提示miR-200a高表达组和miR-200a+SATB1共表达组的SATB1表达较未转染组低,表明miR-200a可以抑制SATB1蛋白表达,亦表明SATB1是miR-200a的靶基因。结论miR-200a在前列腺癌组织中呈低表达,与前�
Objective To explore the expression of microRNA(miRNA,miR)-200a in prostate cancer tissues and its correlation with clinical parameters,and to study the effect of miR-200a on tumor growth in nude mice in vivo.Methods Fluorescence in situ hybridization(FISH)technology was used to detect the expression of miRNA-200a in 54 cases of prostate cancer tissues confirmed by pathology from Xuzhou Medical University from January 2018 to August 2019 Correlation between score,prostate specific antigen(PSA)level,and age;Establish a mouse(purchased from Shanghai Xipuer-Bikai Experimental Animal Co.,Ltd.)subcutaneous transplantation tumor model,divided into three groups:DU145 cell group transfected with miRNA-200a high expression plasmid 2.Transfect the plasmid group of DU145 cells expressing miRNA-200a+SATB1 and the non-transfected DU145 cell group,inoculate 0.2 ml of cell suspension subcutaneously in the right posterior hip of mice,measure the volume of subcutaneous transplanted tumor every 5 days,and draw naked The growth curve of the transplanted tumors in mice was analyzed to analyze the statistical differences in the volume of the transplanted tumors in the three groups.The expression of SATB1 was detected by immunohistochemistry and Western blot.SPSS 25.0 statistical software was used for analysis.The counted data was expressed as a rate(or percentage).The positive rate was compared using theχ^2 test(when the theoretical frequency was less than 5,the Fisher exact probability method was used),and the rank data was compared using the rank sum test and Spearman.Level-related analysis.The Western blotting experiment results were analyzed using Image J grayscale software.Compared with the results of the control protein analysis,the relative protein analysis results showed that the relative gray value was statistically significant with P<0.05 as the difference.The data are expressed as mean±standard deviation(Mean±SD),single-factor analysis of variance is used for comparison between multiple samples,and t-test analysis is us
作者
刘一锐
李柄恒
于海元
马赛
阚懿
杨栋梁
毛立军
Liu Yirui;Li Bingheng;Yu Haiyuan;Ma Sai;Kan Yi;Yang Dongliang;Mao Lijun(Department of Urology,the Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第2期356-359,共4页
Chinese Journal of Experimental Surgery
基金
江苏省青年医学人才基金(QNRC2016794)。
