ABSTRACT
Objection@#To investigate the effect of miR-32-5p on the radiosensitivity, migration and invasion of colorectal cancer cells and the underlying mechanism.@*Methods@#Human colorectal cancer SW480 cells and normal colonic epithelial NCM460 cells were cultured. The colorectal cancer cells were divided into the non-transfected and transfected groups (transfected with anti-miR-NC, anti-miR-32-5p, pcDNA, pcDNA-TOB1, anti-miR-32-5p+ si-NC and anti-miR-32-5p+ si-TOB1, respectively). The expression of miR-32-5p and TOB1 at the mRNA and protein levels was detected by RT-qPCR and Western blot. The radiosensitivity of the transfected cells was determined by colony formation assay. The migration and invasion ability of the transfected cells were detected by Transwell assay. Whether miR-32-5p targeted TOB1 was validated by dual luciferase reporter gene assay and Western blot.@*Results@#Compared with human colonic epithelial cells, the expression of miR-32-5p was significantly up-regulated, whereas the expression of TOB1 mRNA and protein was remarkably down-regulated in the colon cancer cells (all P<0.05). Compared with the anti-miR-NC, the quantity of cell migration and invasion was significantly decreased (both P<0.05) and the radiosensitivity ratio was 1.801 in the anti-miR-32-5p group. Compared with the pcDNA group, the quantity of cell migration and invasion was significantly decreased (both P<0.05) and the radiosensitivity ratio was 1.764 in the pcDNA-TOB1 group. Dual luciferase reporter gene assay and Western blot confirmed that miR-32-5p negatively regulated the expression of TOB1 protein. Compared with the anti-miR-32-5p+ si-NC group, the quantity of cell migration and invasion was significantly increased (both P<0.05) and the radiosensitivity ratio was 0.591 in the anti-miR-32-5p+ si-TOB1 group.@*Conclusions@#Inhibition of miR-32-5p expression can significantly enhance the radiosensitivity of colorectal cancer cells and suppress cell migration and invasion. The underlying mechanism might be related to the targeted up-regulation of TOB1 expression.
ABSTRACT
Objection To investigate the effect of miR-32-5p on the radiosensitivity,migration and invasion of colorectal cancer cells and the underlying mechanism.Methods Human colorectal cancer SW480 cells and normal colonic epithelial NCM460 cells were cultured.The colorectal cancer cells were divided into the non-transfected and transfected groups (transfected with anti-miR-NC,anti-miR-32-5p,pcDNA,pcDNA-TOB1,anti-miR-32-5p+si-NC and anti-miR-32-5p+si-TOB1,respectively).The expression of miR-32-5p and TOB1 at the mRNA and protein levels was detected by RT-qPCR and Western blot.The radiosensitivity of the transfected cells was determined by colony formation assay.The migration and invasion ability of the transfected cells were detected by Transwell assay.Whether miR-32-5p targeted TOB1 was validated by dual luciferase reporter gene assay and Western blot.Results Compared with human colonic epithelial cells,the expression of miR-32-5p was significantly up-regulated,whereas the expression of TOB1 mRNA and protein was remarkably down-regulated in the colon cancer cells (all P＜0.05).Compared with the anti-miR-NC,the quantity of cell migration and invasion was significantly decreased (both P＜0.05) and the radiosensitivity ratio was 1.801 in the anti-miR-32-5p group.Compared with the pcDNA group,the quantity of cell migration and invasion was significantly decreased (both P＜0.05) and the radiosensitivity ratio was 1.764 in the pcDNA-TOB1 group.Dual luciferase reporter gene assay and Western blot confirmed that miR-32-5p negatively regulated the expression of TOB1 protein.Compared with the anti-miR-32-5p+si-NC group,the quantity of cell migration and invasion was significantly increased (both P＜0.05) and the radiosensitivity ratio was 0.591 in the anti-miR-32-5p+si-TOB1 group.Conclusions Inhibition of miR-32-5p expression can significantly enhance the radiosensitivity of colorectal cancer cells and suppress cell migration and invasion.The underlying mechanism might be related to the targeted up-regulation of TOB1 expression.
ABSTRACT
OBJECTIVE:To investigate the effects of SDZ on LOVO cell line and the expression of E-Cadherin and N-Cadherin. METHODS: The apoptosis of SDZ on growth of LOVO cells was observed under electron microscope and inverted microscope. The expression of E-Cadherin and N-Cadherin treated by SDZ were detected by RT-PCR. RESULTS: SDZ promoted the apoptosis of LOVO cell lines and increased the expression of E-Cadherin,however,it did no effect on the expression of N-Cadherin. CONCLUSION: SDZ can enhance the intercellular adhesiveness and degrade the capability of infiltration and metastasis of tumor cells.
ABSTRACT
Objective To study the effect of selective cyclooxygenase-2 inhibitor celecoxib on cell apoptosis of human colorectal cancer cell line HT-29 and the probable mechanism involved by detecting the expressions of cytochrome C, Caspase-9 and poly ADP-ribose polymerase(PARP) at protein level. Methods Apoptosis was determined by Acridine orange and Ethidium bromide staining under fluorescence microscope and flow cytometry. The protein expression of cytochrome C, Capsase-9 and PARP were examined by Western blotting.Results Celecoxib induced apoptosis of HT-29 human colorectal cancer cells in a concentration and time-dependent manner from 0 to 120 ?mol/L. Sub-G 1 peak was detected by flowcytometry, and the apoptotic rate was between(7.31?2.37)%-(48.30 ?2.86)%. Celecoxib induced cytochrome C release into the cytosol from mitochondria, then activated Caspase-9 and consequently triggered PARP cleavage.Conclusion Celecoxib can induce apoptosis through a cytochrome C-dependent pathway in human colorectal cancer cell line HT-29.