miRNA-34a反义寡核苷酸对非小细胞肺癌细胞株HCC827的影响

L. Ren, Yu Wang, S. Pang, L. Fan
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HCC827 cells were divided into three groups: blank control group, negative control group, anti-sense oligonucleotide group(liposome 2000 transfected anti-sense oligonucleotide miRNA-34a); cell counting kit-8 (CCK-8) method was used to detect cell proliferation, Jimsa staining was used to detect cell cloning ability, Transwell test was used to detect cell migration and invasion ability; RT-PCR and Western blot were used to detect phosphatase and tensin homolog (PTEN), phosphorylation-protein kinase B (p-Akt), phosphatidylinositol-3-kinase (PI3K) mRNA and protein expression. \n \n \nResults \nThe relative expression of miRNA34a in HCC827 cells was significantly higher than that in human normal lung cells (P 0.05). At 48 h, 72 h and 96 h, the proliferation level of HCC827 cells in anti-sense oligonucleotide miRNA-34a group was significantly lower than that in negative control group and blank control group (P<0.05). The cell cloning rate of antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group (P<0.01). The number of migration and invasion of HCC827 cells in antisense oligonucleotide RNA-34a group was significantly lower than that in negative control group and blank control group (P<0.01). The relative expression of PTEN mRNA and protein in antisense oligonucleotide miRNA-34a group was significantly higher than that in negative control group and blank control group (P<0.05); the relative expression of p-Akt, PI3K mRNA and protein in antisense oligonucleotide miRNA-34a group were significantly lower than that in negative control group and blank control group (P<0.05). \n \n \nConclusions \nThe expression level of miRNA-34a in human non-small cell lung cancer cells is significantly higher than that in human normal lung cells. 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引用次数: 0

摘要

目的探讨miRNA-34a反义寡核苷酸在非小细胞肺癌(NSCLC)中的作用及其分子机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测人非小细胞肺癌细胞株HCC827和人正常肺细胞MRC-5中miRNA-34a的表达。将HCC827细胞分为3组:空白对照组、阴性对照组、反义寡核苷酸组(脂质体2000转染反义寡核苷酸miRNA-34a);细胞计数试剂盒-8 (CCK-8)法检测细胞增殖,Jimsa染色法检测细胞克隆能力,Transwell试验检测细胞迁移和侵袭能力;采用RT-PCR和Western blot检测磷酸酶和紧张素同源物(PTEN)、磷酸化蛋白激酶B (p-Akt)、磷脂酰肌醇-3激酶(PI3K) mRNA和蛋白的表达。结果miRNA34a在HCC827细胞中的相对表达量显著高于正常肺细胞(P < 0.05)。在48 h、72 h和96 h,反义寡核苷酸miRNA-34a组HCC827细胞的增殖水平显著低于阴性对照组和空白对照组(P<0.05)。反义寡核苷酸miRNA-34a组细胞克隆率显著低于阴性对照组和空白对照组(P<0.01)。反义寡核苷酸RNA-34a组HCC827细胞的迁移和侵袭数量显著低于阴性对照组和空白对照组(P<0.01)。反义寡核苷酸miRNA-34a组PTEN mRNA和蛋白相对表达量显著高于阴性对照组和空白对照组(P<0.05);反义寡核苷酸miRNA-34a组P - akt、PI3K mRNA和蛋白的相对表达量显著低于阴性对照组和空白对照组(P<0.05)。结论miRNA-34a在人非小细胞肺癌细胞中的表达水平明显高于人正常肺细胞。miRNA-34a反义寡核苷酸可抑制人非小细胞肺癌细胞的增殖、克隆、迁移和侵袭。其机制可能与PTEN/p-Akt/PI3K信号通路的负调控有关。关键词:寡核苷酸;反义;miRNA-34a;肺癌,非小细胞肺;细胞系、肿瘤
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The effect of miRNA-34a antisense oligonucleotide on non-small cell lung cancer cell line HCC827
Objective To investigate the effect of antisense oligonucleotides of miRNA-34a on non-small cell lung cancer (NSCLC) and its molecular mechanism. Methods The expression of miRNA-34a in human non-small cell lung cancer cell line HCC827 and human normal lung cell MRC-5 was detected by real time fluorescence quantitative polymerase chain reaction (qRT-PCR). HCC827 cells were divided into three groups: blank control group, negative control group, anti-sense oligonucleotide group(liposome 2000 transfected anti-sense oligonucleotide miRNA-34a); cell counting kit-8 (CCK-8) method was used to detect cell proliferation, Jimsa staining was used to detect cell cloning ability, Transwell test was used to detect cell migration and invasion ability; RT-PCR and Western blot were used to detect phosphatase and tensin homolog (PTEN), phosphorylation-protein kinase B (p-Akt), phosphatidylinositol-3-kinase (PI3K) mRNA and protein expression. Results The relative expression of miRNA34a in HCC827 cells was significantly higher than that in human normal lung cells (P 0.05). At 48 h, 72 h and 96 h, the proliferation level of HCC827 cells in anti-sense oligonucleotide miRNA-34a group was significantly lower than that in negative control group and blank control group (P<0.05). The cell cloning rate of antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group (P<0.01). The number of migration and invasion of HCC827 cells in antisense oligonucleotide RNA-34a group was significantly lower than that in negative control group and blank control group (P<0.01). The relative expression of PTEN mRNA and protein in antisense oligonucleotide miRNA-34a group was significantly higher than that in negative control group and blank control group (P<0.05); the relative expression of p-Akt, PI3K mRNA and protein in antisense oligonucleotide miRNA-34a group were significantly lower than that in negative control group and blank control group (P<0.05). Conclusions The expression level of miRNA-34a in human non-small cell lung cancer cells is significantly higher than that in human normal lung cells. Antisense oligonucleotides of miRNA-34a can inhibit the proliferation, cloning, migration and invasion of human non-small cell lung cancer cells. The mechanism may be related to the negative regulation of PTEN/p-Akt/PI3K signaling pathway. Key words: Oligonucleotides, antisense; miRNA-34a; Carcinoma, non-small-cell lung; Cell line, tumor
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中国医师杂志
中国医师杂志 Medicine-Medicine (all)
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