ABSTRACT
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 miRNA34a 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 ＜ 0.01).The relative expression of miRNA34a in antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group (P ＜ 0.05),and there was no significant difference between negative control group and blank control group (P ＞ 0.05).At 48 h,72 h and 96 h,the proliferation level of HCC827 cells in antisense 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 nonsmall 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.
ABSTRACT
Purpose This subject detected the expression of paper miRNA34a,c-myc gene and protein in adenocarcinoma of lung tissue,adjacent mucosa tissue and normal lung tissue exploring the relationship between the two and the clinical significance.Method The project mainly used the method of real-time fluorescent quantitative PCR (qRT-PCR),Western blot and immunohistochecical to detect the expression of miRNA34a and c-myc in normal tissue and adenocarcinoma of lung respectively.Result (1) lower expression of miRNA34a in lung adenocarcinoma tissue,lower than the adjacent and normal tissues (P ＜ 0.01),higher than the metastatic carcinoma group (P ＜ 0.01).C-myc is highly expressed in lung adenocarcinoma tissue,higher than the adjacent and normal tissues (P ＜0.01),lower than the metastatic carcinoma tissue (P ＜ 0.01).miRNA34a and c-myc beside carcinoma tissues and normal tissues also have significant difference between the statistical significance (P ＜ 0.05).(2) Differentiation of low and high there was no statistically significant difference between patients with lung adenocarcinoma (P ＞ 0.05).there were significant differences in the expression of miRNA34a and c-myc in the tissues of metastatic and non metastatic adnocarcinoma.(3)The positive expression of c-myc group of lung cancer patients was lower than that of the negative group (P ＜ 0.05),low expression of the survival time in miRNA34a group was significantly lower than that in high expression group,there was significant difference (P ＜ 0.01).Conclusion Expression of miRNA34a and c-myc both showed a negative correlation in adenocarcinoma of lung.
ABSTRACT
Objective To detect the expression of miRNA -34a in normal cervical,low grade cervical intra-epithelial neoplasia and high grade cervical intraepithelial neoplasia,to explore its clinical significance.Methods The expression levels of miRNA -34a in 82 cases of low grade cervical intraepithelial neoplasia (CINⅠ,CIN L group),87 cases of high grade cervical intraepithelial neoplasia (CINⅡ -Ⅲ,CIN H group) and 60 cases of normal cervical tissues (control group) in tissue specimens and serum were examined by real -time PCR(RT -PCR). Results The relative expression levels of miRNA -34a in tissues and serum of CIN H group were (0.584 ± 0.288),(0.528 ±0.299),which of CIN L group were (0.903 ±0.252),(0.847 ±0.279),which of the control group were (0.960 ±0.212),(0.908 ±0.223).The miRNA -34a level of CIN H group in tissues and serum were lower than CIN L group and control group,there was significant difference( Ptissues =0.001,Pserum =0.003).The miRNA -34a level in tissue and serum had no significant difference between the CINL group and control group(P >0.05).The expression level of serum miRNA -34a was positively related with the level of the tissues(r =0.907,P <0.01).Conclusion The expression level of miRNA -34a in high grade CIN was significantly decreased,which indi-cated that miRNA -34a play an important role in the prognosis of CIN and the transformation of cervical cancer,and it has certain clinical value for the diagnosis and treatment of CIN.