摘要
目的探讨微小RNA-216a(miR-216a)对宫颈癌细胞增殖、凋亡的影响及作用机制。方法收集2019年5月至2021年5月于河北北方学院附属第一医院行手术治疗的82例宫颈癌患者的宫颈癌组织和癌旁正常组织,用实时荧光定量PCR(qRT-RCR)法检测组织中miR-216a和钙激活中性蛋白酶6(CAPN6)表达。转染miR-216a模拟物(miR-216a mimic)、CAPN6小干扰RNA(si-CAPN6)或共转染miR-216a mimic与CAPN6过表达质粒(pcDNA-CAPN6)至宫颈癌HeLa细胞。采用细胞计数试剂盒(CCK-8)法检测细胞增殖,流式细胞术检测细胞凋亡,蛋白质印迹法检测CAPN6、切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved Caspase-3)、B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)的蛋白表达情况,并进行统计分析。结果宫颈癌组织中miR-216a mRNA相对表达量明显低于癌旁正常组织(t=52.968,P<0.05),而CAPN6 mRNA和CAPN6蛋白相对表达量均明显高于癌旁正常组织(t值分别为26.622、45.036,P<0.05)。过表达miR-216a降低了HeLa细胞24、48、72h的增殖活力及Bcl-2蛋白相对表达量,提高了细胞凋亡率及Cleaved Caspase-3、Bax蛋白相对表达量(F=11.885~90.813,P<0.05)。过表达miR-216a降低了HeLa细胞中CAPN6 mRNA和CAPN6蛋白相对表达量(F值分别为58.990、156.82,P<0.05)。下调CAPN6降低了HeLa细胞24、48、72h的增殖活力及Bcl-2蛋白相对表达量,提高了细胞凋亡率及Cleaved Caspase-3、Bax蛋白相对表达量(F=42.390~241.912,P<0.05)。过表达CAPN6可逆转过表达miR-216a对HeLa细胞增殖、凋亡及凋亡蛋白的影响。结论miR-216a在宫颈癌组织中低表达,过表达miR-216a可抑制宫颈癌细胞增殖并促进细胞凋亡,其可能是通过靶向下调CAPN6表达发挥作用。
Objective To investigate the effect of microRNA-216 a(miR-216 a)on the proliferation and apoptosis of cervical cancer cells and its mechanism.Methods The cervical cancer tissues and normal adjacent tissues of 82 patients with cervical cancer who underwent surgical in the First Affiliated Hospital of Hebei North University from May 2019 to May 2021 were collected.The expressions of miR-216 a and calcium activated neutral protease 6(CAPN6)in the tissues were detected by real-time quantitative reverse transcriptase polymerase chain reaction(qRT-RCR).The miR-216 a simulant(miR-216 a mimic),CAPN6 small interfering RNA(si-CAPN6)or miR-216 a mimic and CAPN6 overexpression plasmid(pcDNA-CAPN6)were transfected to cervical cancer HeLa cells.Cell proliferation was detected by cell counting kit-8(CCK-8),apoptosis was detected by flow cytometry.The protein expressions of CAPN6,cleaved caspase-3,B cell CLL/lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)were detected by Western blot and analyzed statistically.Results The relative expression level of miR-216 a mRNA in cervical cancer was significantly lower than that in adjacent normal tissues(t=52.968,P<0.05),while the expression level of CAPN6 mRNA and CAPN6 protein was significantly higher than those in adjacent normal tissues(t=26.622 and 45.036,respectively,P<0.05).The overexpression of miR-216 a decreased the proliferation activity of HeLa cells at 24,48 and 72 hours and Bcl-2 protein expression,and increased the apoptosis rate and the expression of cleaved caspase-3 and Bax protein(F=11.885-90.813,P<0.05).The overexpression of miR-216 a significantly decreased the relative expression level of CAPN6 mRNA and CAPN6 protein in HeLa cells(F=58.990 and 156.82,respectively,P<0.05).The down regulation of CAPN6 decreased the proliferation activity of HeLa cells at 24,48 and 72 hours and Bcl-2 protein expression,and increased the apoptosis rate and the expression of cleaved caspase-3 and Bax protein(F=42.390-241.912,P<0.05).The overexpression of CAPN6 could reverse the effect
作者
张贤雨
马欢
宋凤丽
张志林
ZHANG Xianyu;MA Huan;SONG Fengli;ZHANG Zhilin(Department of Radiotherapy,The First Affiliated Hospital of Hebei North University,Hebei Zhangjiakou 075000,China;Department of Radiotherapy,The Third Affiliated Hospital of Beijing University of Traditional Chinese Medicine,Beijing 100029,China)
出处
《中国妇幼健康研究》
2023年第1期75-84,共10页
Chinese Journal of Woman and Child Health Research
基金
张家口市市级科技计划资助项目(2121192H)。
