Dyrk1b蛋白在不同程度子宫颈病变组织及子宫颈癌细胞中的表达及意义  

Expression and clinical significance of Dyrklb in the specimens and cells of cervical lesions

作　　者：林姣[1] 张朋新[2] 庞露[1] 陈婧盈 孙雯[1] 祁文静[2] 吕恽怡[1] 关宏伟[2] 高景春[1] 

机构地区：[1]大连医科大学附属第一医院妇产科,116011 [2]大连医科大学附属第一医院病理科,116011

出　　处：《中华妇产科杂志》2016年第1期40-45,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：辽宁省科技计划（2012225020）

摘　　要：目的探讨双特异性酪氨酸磷酸化调节激酶1b（Dyrk1b）蛋白在不同程度子宫颈病变组织及子宫颈癌细胞中的表达及意义。方法（1）组织标本检测：选取2011年1月至2013年12月间就诊于大连医科大学附属第一医院，经病理检查证实的子宫颈鳞癌75例（其中I期60例，Ⅱ期15例）、低级别鳞状上皮内病变（LSIL）12例、高级别鳞状上皮内病变（HSIL）40例患者的石蜡组织标本；另取同期经病理检查确诊的28例慢性子宫颈炎患者的子宫颈组织为对照。采用免疫组化SP法检测不同程度子宫颈病变组织中Dyrk1b蛋白的表达情况，并分析子宫颈癌组织中Dyrk1b蛋白的表达与其不同临床病理指标的关系。（2）细胞实验：采用子宫颈癌细胞系HeLa和SiHa细胞，实验分为两组，实验组细胞中加入Dyrk1b抑制剂——AZ191，对照组细胞中不加AZ191。采用蛋白印迹法检测不同浓度（5、10μmol／L）的AZ191作用后两组细胞中Dyrk1b蛋白的表达；四甲基偶氮唑蓝（MTT）比色法检测不同浓度（2．5、5、10、25、50、100μmol／L）的AZ191作用后两组细胞的增殖情况；流式细胞仪检测不同浓度（5、10μmol／L）的AZ191作用后两组细胞的凋亡情况。结果（1）免疫组化sP法检测显示，慢性子宫颈炎、LSIL、HSIL和子宫颈鳞癌组织中Dyrk1b蛋白阳性表达率分别为11％（3／28）、1／12、42％（17／40）、71％（53／75），子宫颈鳞癌明显高于慢性子宫颈炎、LSIL、HSIL（P〈0．01）；HSIL明显高于慢性子宫颈炎、LSIL（P〈0．05）；而LSIL号熳性子宫颈炎比较，差异无统计学意义（P〉0．05）。子宫颈鳞癌组织中Dyrk1b蛋白的表达与子宫颈肌层浸润深度显著相关（P〈0．01），而与年龄、临床分期、淋巴结有无转移以及血清鳞状上皮细胞癌抗原（SCC—Ag）水平均无显著相关性（P〉0．05）。（2）蛋白印迹法检测显示，不同浓度（5、10μmoObjective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinaselb （Dyrklb） in the specimens and ceils of cervical lesions. Methods （ 1 ） All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion （SIL） admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stage I and 15 cases of stage I1, 12 cases with low-grade squamous intraepithelial lesion （LSIL）and 40 cases with high- grade sqnamous intraepithelial lesion （HSIL）. While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrklb was detected by immunohistochemistry among the four groups. （2） The expression of Dyrklb in HeLa and SiHa cells were detected by western blot method and the expression of Dyrklb protein were also detected after treatment of AZ191 （5, 10 μmol/L） for 48 hours in HeLa and SiHa cells. （3）The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191 （2.5, 5, 10, 25, 50, 100 μmol/L）for 48 hours were detected by methyl thiazolyl tetrazolium （MTT） assay. （4） The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZI91 （5, 10 μmol/L） for 48 hours. Results （1） The positive rates of Dyrklb protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%（3/28）, 1/12, 42% （17/40） and 71% （53/75）, respectively. The expression of Dyrklb in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis （P 〈 0.01）, there were significant difference between cervical sqnamous cancer and HSIL, or between HSIL and LSIL（all P 〈 0.05）, while there were not significant difference between LSIL and chronic cervicitis （P 〉 0.05）. Expression of Dyrklb was corre

关 键 词：蛋白酪氨酸激酶类 蛋白质丝氨酸苏氨酸激酶 宫颈肿瘤 宫颈上皮内瘤样病变 细胞凋亡 印迹法 蛋白质 

分 类 号：R737.33[医药卫生—肿瘤]