| Background : According to statistics,among 185 countries in the world,colorectal cancer is a malignant tumor with the third incidence rate and the second mortality rate.The incidence and mortality of colorectal cancer have increased year by year in China in recent 30 years with the rapid development of economy and the change of people’s lifestyle.Micro RNAs(miRNAs)can participate in the occurrence and development of a variety of tumors by regulating related genes at the post-transcriptional level,and with complex mechanisms.Autophagy is a highly conservative circulatory process for cell survival and maintenance of steady state.Currently,it has been found that a variety of miRNA can participate in tumor progression through autophagy.As one of them,miR-186-5p has been proved to play different roles in different cancer species by previous studies,which can promote or inhibit cancer.However,its role in colorectal cancer has not yet been clarified.Purpose:To confirm the expression of Micro RNA-186-5p(miR-186-5p)is significantly different in normal cancer,cancer cells,normal tissues and cancer tissues,and it is further confirmed that miR-186-5p can affect the related biological functions of colorectal cancer,analyze its correlation with autophagy-related factors and clinicopathological features of patients with colorectal cancer,and find new targets that can predict and treat colorectal cancer.Materials and methods:1)The colorectal normal intestinal epithelial cell line(NCM460 cells)and cancer cell lines(RKO cells,LOVO cells)were cultured.The cells with different expression levels of miR-186-5p compared with normal intestinal epithelial cells were screened by q RT-PCR for subsequent experiments,and the relative expression levels of autophagy-related factors in colorectal cancer cells were detected compared with normal intestinal epithelial cells.2)LOVO cell line was selected,and miR-186-5p inhibitor was used to transfect cancer cells with good growth status in logarithmic phase.q RT-PCR were used to detect the expression levels of autophagy-related factors(p62,Beclin-1,LC3-II).CCK-8 and Wound-Healing Assay were used to detect the changes of cell proliferation and migration biological functions.3)Forty cases of postoperative tissue specimens of colorectal cancer from 2016 to2022 in the Affiliated Hospital of Yan ’an University were collected(all clinicopathological data were complete).q RT-PCR were used to detect the expression of miR-186-5p in normal tissues and cancer tissues.Statistical analysis was performed to analyze whether there was a significant difference between normal and cancer tissues and to analyze whether the expression of miR-186-5p is related to the clinical and pathological factors of patients.Results:1)Compared with normal intestinal epithelial cells,the expression of miR-186-5p in LOVO cells is relatively high,and the difference is statistically significant(p < 0.05).Compared with normal intestinal epithelial cells,the expression levels of autophagy-related factors Beclin-1 and LC3-II in LOVO cells were higher,and the expression level of P62 was lower,with statistical significance(p < 0.05);2)Compared with NC group,the expression of autophagy related factors Beclin-1and LC3-II in LOVO cells transfected with miR-186-5p inhibitor decreased,and the difference was not statistically significant(p > 0.05),while the expression level of p62 increased significantly,with statistical significance(p < 0.05);3)The LOVO cells transfected with miR-186-5p inhibitor were used for scratch experiment.The results showed that the scratch healing of LOVO cells after transfection was slower than that of NC group,and the difference between them was statistically significant after quantitative comparison(p < 0.05);4)CCK-8 experiment was carried out on LOVO cells transfected with miR-186-5p inhibitor.The results showed that compared with the NC group,the proliferation of LOVO cells after transfection was slower,and the difference was statistically significant(P < 0.05).5)The expression of miR-186-5p was significantly different between normal colorectal tissues and cancer tissues(p < 0.05);6)The expression of miR-186-5p was not correlated with age,sex,tumor location,type,size,pathological type,differentiation degree,neurovascular invasion and lymph node metastasis(p > 0.05);Conclusion:1)The expression of miR-186-5p was up-regulated in colorectal cancer cells and tissues,it promotes the proliferation,migration and invasion of colorectal cancer cells and affects the expression of autophagy-associated factor p62 in colorectal cancer cells;2)The expression of miR-186-5p was not correlated with age,sex,tumor location,histological type,presence or absence of neurovascular invasion,tumor size,gross type,depth of invasion,degree of differentiation,and lymph node metastasis. |
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