首页|基因间长链非编码RNA467对子宫内膜癌细胞增殖、凋亡及迁移和侵袭能力的影响

基因间长链非编码RNA467对子宫内膜癌细胞增殖、凋亡及迁移和侵袭能力的影响

Effects of long intergenic non-coding RNA 467 on proliferation,apoptosis and migration,invasion ability of endometrial carcinoma cells

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目的 探讨基因间长链非编码RNA 467(linc00467)对子宫内膜癌细胞增殖、凋亡及迁移和侵袭能力的影响.方法 体外培养子宫内膜癌细胞株HEC-1A、Ishikawa、KLE和RL-95-2,应用实时荧光定量聚合酶链反应(RT-qPCR)法检测4种细胞中linc00467的相对表达量,选择高表达linc00467的子宫内膜癌细胞株HEC-1A、Ishikawa进行后续实验.分别构建2个linc00467慢病毒沉默表达载体sh-linc00467#1、sh-linc00467#2和空载慢病毒质粒;将对数生长期HEC-1A、Ishikawa细胞分为sh-NC组、sh-linc00467#1组和sh-linc00467#2组,sh-NC组细胞转染空载慢病毒质粒,sh-linc00467#1 组和 sh-linc00467#2 组细胞分别转染 sh-linc00467#1 和 sh-linc00467#2;应用 RT-qPCR 法检测 3 组细胞中linc00467的相对表达量,5-乙炔基-2-脱氧尿苷(EdU)实验、克隆形成实验检测3组细胞增殖能力,流式细胞术检测3组细胞凋亡情况,划痕实验检测3组细胞的迁移能力,Transwell小室实验检测3组细胞侵袭能力.结果 HEC-1A细胞中linc00467 mRNA的相对表达量显著高于KLE和RL-95-2细胞(P<0.05);Ishikawa细胞中linc00467 mRNA的相对表达量显著高于KLE和RL-95-2细胞(P<0.05);HEC-1A与Ishikawa细胞中linc00467 mRNA的相对表达量比较差异无统计学意义(P>0.05),KLE细胞中linc00467 mRNA的相对表达量显著低于RL-95-2细胞(P<0.05).sh-linc00467#1组和 sh-linc00467#2 组 HEC-1A、Ishikawa 细胞中 linc00467 mRNA 的相对表达量显著低于 sh-NC组(P<0.01);sh-linc00467#1 组与 sh-linc00467#2 组 HEC-1A、Ishikawa 细胞中 linc00467 mRNA 的相对表达量比较差异无统计学意义(P>0.05).sh-linc00467#1组和sh-linc00467 #2组EdU阳性HEC-1A、Ishikawa细胞数显著少于sh-NC 组(P<0.01);sh-linc00467#1 组与 sh-linc00467#2 组 EdU 阳性 HEC-1A、Ishikawa 细胞数比较差异无统计学意义(P>0.05).sh-linc00467#1 组和 sh-linc00467#2 组 HEC-1A、Ishikawa 细胞克隆数显著少于 sh-NC 组(P<0.01);sh-linc00467#1 组与 sh-linc00467#2 组 HEC-1 A、Ishikawa 细胞克隆数比较差异无统计学意义(P>0.05).sh-linc00467#1组和 sh-linc00467#2 组 HEC-1 A、Ishikawa 细胞凋亡率显著高于 sh-NC 组(P<0.01);sh-linc00467#1 组与 sh-linc00467#2组 HEC-1 A、Ishikawa 细胞凋亡率比较差异无统计学意义(P>0.05).sh-linc00467#1 组和 sh-linc00467#2 组 HEC-1A、Ishikawa 细胞划痕愈合率显著低于 sh-NC 组(P<0.01);sh-linc00467#1 组与 sh-linc00467#2 组 HEC-1 A、Ishikawa 细胞划痕愈合率比较差异无统计学意义(P>0.05).sh-linc00467#1组和sh-linc00467#2组HEC-1 A、Ishikawa侵袭细胞数显著少于sh-NC组(P<0.01);sh-linc00467#1组与sh-linc00467#2组HEC-1 A、Ishikawa侵袭细胞数比较差异无统计学意义(P>0.05).结论 下调linc00467表达可抑制子宫内膜癌细胞的增殖、迁移和侵袭,促进子宫内膜癌细胞凋亡.
Objective To investigate the effects of long intergenic non-coding RNA 467(linc00467)on the proliferation,apoptosis and migration,invasion ability of endometrial carcinoma cells.Methods Human endometrial carcinoma cells HC1A,Ishikawa,KLE and RL-95-2 were cultured in vitro,the expression level of linc00467 in the four cells was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Endometrial carcinoma cell lines HEC-1A and Ishikawa with the highest linc00467 expression levels were selected for subsequent experiment.Two Iinc00467 lentivirus silencing expression vectors of sh-linc00467#1 and sh-linc00467#2,and empty lentivirus plasmids were constructed,respectively;the HEC-1A and Ishikawa cells in logarithmic growth phase were divided into sh-NC group,sh-linc00467#1 group,and sh-linc00467#2 group.The cells in the sh-NC group were transfected with empty lentivirus plasmids,the cells in the sh-linc00467#1 group and sh-linc00467#2 group were transfected with sh-linc00467#1 and sh-linc00467#2,respectively;the relative expression level of linc00467 in cells of the three groups was detected by RT-qPCR,the proliferation ability of cells in the three groups was detected by 5-ethynyl-2-deoxyuridine(EdU)assay and colony formation assay,the migration ability of cells in the three groups was detected by scratch assay,the invasion ability of cells in the three groups was detected by Transwell assay,and the cell apoptosis in the three groups was detected by flow cytometry.Results The relative expression level of linc00467 mRNA in HEC-1A cells was significantly higher than that in KLE and RL-95-2 cells(P<0.05);the relative expression level of linc00467 mRNA in Ishikawa cells was significantly higher than that in KLE and RL-95-2 cells(P<0.05);there was no statistically significant difference in the relative expression level of linc00467 mRNA between HEC-1A and Ishikawa cells(P>0.05);the relative expression level of linc00467 mRNA in KLE cells was significantly lower than that in RL-95-2 cells(P<0.05).The relative expression levels of linc00467 mRNA in HEC-1A and Ishikawa cells in the sh-linc00467#1 group and sh-linc00467#2 group were significantly lower than those in the sh-NC group(P<0.01);there was no statistically significant difference in the relative expression level of linc00467 mRNA in HEC-1A and Ishikawa cells between the sh-linc00467 # 1 group and the sh-linc00467#2 group(P>0.05).The number of EdU positive HEC-1A and Ishikawa cells in the sh-linc00467#1 group and sh-linc00467#2 group was significantly lower than that in the sh-NC group(P<0.01);there was no statistically significant difference in the number of EdU positive HEC-1A and Ishikawa cells between the sh-linc00467#1 group and sh-linc00467#2 group(P>0.05).The number of cloned HEC-1A and Ishikawa cells in the sh-linc00467#1 group and sh-linc00467#2 group was significantly lower than that in the sh-NC group(P<0.01);there was no statistically significant difference in the number of cloned HEC-1A and Ishikawa cells between the sh-linc00467#1 group and sh-linc00467#2 group(P>0.05).The apoptosis rates of HEC-1A and Ishikawa cells in the sh-linc00467#1 group and sh-linc00467#2 group were significantly higher than that in the sh-NC group(P<0.01);there was no statistically significant difference in the apoptosis rates of HEC-1A and Ishikawa cells between the sh-linc00467#1 group and sh-linc00467 #2 group(P>0.05).The scratch healing rates of HEC-1A and Ishikawa cells in the sh-linc00467#1 group and sh-linc00467#2 group were significantly lower than those in the sh-NC group(P<0.01);there was no statistically significant difference in the scratch healing rates of HEC-1 A and Ishikawa cells between the sh-linc00467#1 group and sh-linc00467#2 group(P>0.05).The number of invasive cells of HEC-1 A and Ishikawa in the sh-linc00467#1 group and sh-linc00467#2 group was significantly lower than that in the sh-NC group(P<0.01);there was no statistically significant difference in the number of invasive cells of HEC-1 A and Ishikawa between the sh-linc00467#1 group and sh-linc00467#2 group(P>0.05).Conclusion Down-regulation of linc00467 expression can inhibit the proliferation,migration and invasion,and promote apoptosis of endometrial carcinoma cells.

endometrial carcinomalong intergenic non-coding RNA 467proliferationmigrationinvasionapoptosis

王圆圆、关新垒、秦海霞

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新乡医学院第一附属医院妇科,河南 卫辉 453100

子宫内膜癌细胞 基因间长链非编码RNA 467 增殖 迁移 侵袭 凋亡

河南省高等学校重点科研项目

19A320030

2024

新乡医学院学报
新乡医学院

新乡医学院学报

CSTPCD
影响因子:0.999
ISSN:1004-7239
年,卷(期):2024.41(1)
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