| Lung cancer is one of the highest malignant tumors of morbidity and mortality in China,of which 85% belongs to non-small cell lung cancer(NSCLC).Due to the onset of concealment,most patients when treatment is already late,even though accompanied by continuous improvement of treatment means,NSCLC patients with 5-year survival rate is still very low.Revealing the development mechanism of NSCLC and prolonging the survival of patients has been a hot topic in malignant tumor research.miRNA is a non-coding small molecule RNA with a length of 21-25 nucleotides.miRNA,which can hybridize to 3'-UTR of the target mRNA sequence,inhibits the expression of the target gene protein and plays a regulatory role,which in turn affects the cell proliferation,differentiation and migration and a series of biological activities.Studies have shown that mi RNAs are associated with the development of multiple malignancies,which act as cancer genes and / or tumor suppressor genes in tumor regulation.A large number of studies have confirmed that mi RNAs play an important role in the development and progression of NSCLC.Objective The current study shows that miRNA have different degrees of abnormal expression in a variety of malignant tumors.The purpose of this study is to explore the mi RNA related non-small cell lung cancer,whether there is abnormal expression,such as mi R-195?miR-145?miR-140-3p?mi R-191?miR-126.The expression of miRNA in NSCLC was selected to analyze the relationship between the expression and the clinical pathological features and the influence on the overall survival of the patients;in vitro study of related miRNA(miR-191)influence the migration and invasion of A549 lung adenocarcinoma cells.Methods Part one: Referring to relevant literature,we identified miR-195,miR-145,miR-140-3p,mi R-191,miR-126,that may be related with NSCLC as the research object.Real-time fluorescence relative quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miRNAs in cancer tissues and adjacent tissues of patients with non-small cell lung cancer.The correlation between the expression level and non small cell lung cancer was analyzed.According to the results of the analysis,the miRNA which might be related to the occurrence and development of non-small cell lung cancer was screened.According to the screening results,the results of the preliminary screening of the samples were expanded,and the clinicopathological features of the patients were collected and the survival status of the patients was followed up to further analyze their correlation with non-small cell lung cancer.Part two: Based on the results of the first step,we determined that miR-191 is the object of our further study.The miRNA-NC and miR-191 inhibitors were transfected into A549 cells by transient transfection.The results were divided into three groups: blank group(Mock),negative control group(NC),experimental group(miR-191 inhibitor).Test the three groups by following:(1)the migration ability of A549 cells was tested by wound healing assay;(2)the incursion ability of A549 cells was tested by transwell assay.Results Part one : In the preliminary screening,we examined the expression of mi R-195,miR-145 and mi R-140-3p in the tumor tissue of 34 cases of NSCLC patients and the corresponding adjacent tissues by RT-qPCR.There was no significant change in the expression of tissue(p>0.05),while compared with tumor tissue,miR-191 and miR-126 were different(p<0.05).Expanded test sample,we found that there are still differences in trends with respect to adjacent tissues of expression,miR-191 expression in tumor tissue increase(p<0.05);the expression of mi R-126 down-regulated in tumor tissues(p < 0.05).Collect the patient's clinical and pathological characteristics of information,telephone follow-up patient survival analysis and found: miR-191 expression increased in patients with low differentiation cancer or lymph node metastasis(p<0.05),and the patients with higher miR-191 expression exhibited worse prognosis(p<0.05);but in miR-126 did not find this difference(p>0.05).Part two: After screening for RT-qPCR and related data analysis,we selected miR-191 as our study.The miRNA-NC and miR-191 inhibitors were successfully transferred into A549 cells by transient transfection and divided into three groups according to the results: blank group(Mock),negative control group(NC),experimental group(miR-191 inhibitor): compared with the Mock group and NC group,the expression level of mi R-191 in the experimental group was decreased by about 98%,the difference was statistically significant(p<0.05);while the expression of miR-191 in the Mock and NC groups was not significantly different(p>0.05).After successful silencing miR-191,we used scratch test to detect the migration ability of A549 cells.Compared with blank control(mock)and negative control(miR-NC),after miR-191 silencing,the distance of the healing of A549 cells was significantly shortened and the healing time was longer,the difference was statistically significant(p<0.05).Transwell assay was used to detect the invasion ability of A549 cells,compared with the mock group(124.00+6.56),miR-NC group(123.00+9.95),miR-191 inhibitor group(38.33+5.69)the number of cells across the membrane decreased significantly,cell invasion ability weakened,the difference was statistically significant(p<0.05).Conclusion 1?miRNA plays an important role in the development of non-small cell lung cancer.miR-191 was up-regulated in patients with non-small cell lung cancer,and increased in patients with low differentiation cancer or lymph node metastasis,and the patients with higher miR-191 expression exhibited worse prognosis;but in miR-126 did not find this difference.2?After miR-191 silencing,the migration ability and the invasive ability of A549 cells was weakened.miR-191 is an oncogene in non-small cell lung cancer,mi R-191 is a cancer gene in non-small cell lung cancer,which can promote the development of the disease by increasing the migration and invasion ability of tumor cells. |
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