Effect Of μ Receptor On Epithelial-mesenchymal Transition,Migration And Invasion Of Colorectal Cancer Cells

Objective:To explore the effects of μ opioid receptor on epithelial-mesenchymal transition;,migration and invasion of colorectal cancer cells.Methods:(1)Immunohistochemical method was used to analyze 30 cases of colorectal cancer and adjacent tissues of gastrointestinal surgery patients in the Second Affiliated Hospital of Kunming Medical University.Western blot(Western-Blot)was used to detect the expression of μ opioid receptors in colorectal cancer and adjacent tissues.(2)Three colorectal cancer cells SW620,SW480 and RKO were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.The μ opioid receptors in the colorectal cancer cell lines SW620,SW480 and RKO were localized by immunofluorescence staining.Cell lines with the highest μ opioid receptor expression levels were screened by immunoblotting.This cell was transfected with the following liposomes:liposomes containing different interference sequences(OPRM ShRNA-1,OPRM ShRNA-2,OPRM ShRNA-3,OPRM ShRNA-4)and negative control lipids body.The interference sequences with the highest silencing efficiency of μ opioid receptors in the four interfering sequences were screened by immunoblotting and subsequent experiments were performed:immunoblotting was used to detect epithelial-mesenchymal transition in cell lines selected before and after lipofection.Indicator protein:changes in expression levels of E-cadherin(E-cadherin),Vimentin(vimentin),Snail(snail protein),N-cadherin(N-cadherin);detection of liposomes by scratch healing assay The migration ability of selected cell lines before and after transfection was used;Transwell cell invasion assay was used to detect the invasive ability of selected cell lines before and after lipofection.Results:(1)The mean staining integral and gray value of μ receptor in colorectal cancer tissues were higher than those in colorectal cancer tissues(P<0.05)·(2)μ opioid receptors are expressed in the nucleus and cytoplasm of colorectal cancer cell lines SW620,SW480,and RKO,but are not expressed in the cell membrane.Theμ opioid receptor has the highest expression level in colorectal cancer cell line SW620.The expression efficiency of p opioid receptor in OPRM ShRNA-1,OPRM ShRNA-3 and OPRM ShRNA-4 sequence silenced colorectal cancer cell line SW620 was lower than that of OPRM ShRNA-2 sequence(P<0.05).Compared with the negative control group,the expression level of E-cadherin in the colorectal cancer cell line SW620 with silencing μ opioid receptor increased(P<0.05),while the expression levels of Vimentin,Snail and N-cadherin decreased(P<0.05).The migration ability of the transfected colorectal cancer cell line SW620 was weaker than that of the negative control group(P<0.05).The invasive ability of the transfected colorectal cancer cell line SW620 was weaker than that of the negative control(P<0.05).Conclusion:(1)The expression level of μ opioid receptor in colorectal cancer tissues is higher than that in adjacent tissues.(2)Silencing μ The expression of opioid receptors not only inhibits epithelial mesenchymal transition of colorectal cancer cells,but also inhibits the migration and invasion of colorectal cancer cells.