ABSTRACT
Objective To investigate the expression of MEX3A in colorectal cancer (CRC), and to explore the effect and mechanism of MEX3A on the proliferation and migration of colorectal cancer cells. Methods Totally 327 cases of data(41 normal tissues and 286 tumor tissues) were obtained from TCGA database, and 104 cases of clinical samples (77 cases tissues and 27 paracancerous tissues) were collected for immunohistochemistry, then analysed the differences in MEX3A expression between CRC tissues and normal tissues. Western blotting and immunofluorescence staining were used to evaluate the differential expression of MEX3A in CRC cell lines. CL187 cells were selected as the follow-up research vector. Small interfering RNA of MEX3A(siMEX3A) was transfected into CL187 cells to inhibit the expression of MEX3A. The proliferation and migration of CL187 cells were measured by MTT, colony formation assay and Transwell assay. The expression of PI3K, p-PI3K, Akt and p-Akt were detected by Western blotting. Results TCGA database, immunohistochemistry and Western blotting analysis showed that MEX3A was highly expressed in colorectal cancer. The result of immunofluorescence staining showed that MEX3A was concentrated in the cytoplasm and the nucleus. In MTT, colony formation assay and Transwell assay, the proliferation and migration ability of CL187 cells in siMEX3A group decreased significantly than those in control group (P<0.05). Western blotting result showed that the expression of p-PI3K and p-Akt in siMEX3A group down-regulated significantly (P<0.05), and the inhibition of proliferation and migration ability of CL187 cells induced by siMEX3A group could be reversed by 740 Y-P via activating the PI3K/Akt signaling pathway. Conclusion MEX3A is highly expressed in colorectal cancer and promotes the proliferation and migration of CRC cells via PI3K/Akt signaling pathway.
ABSTRACT
Objective: To investigate the effects of S100A6 gene silencing on the proliferation and migration of Eca109 human esophagus cancer cells. Methods: The shRNA expression vectors were constructed using the shRNA sequences designed based on human S100A6's coding sequence, and were transfected into Eca109 cells via cationic liposome. The changes of S100A6 mRNA and protein in Eca109 cells transfected with the recombinant vectors were detected using real-time PCR and Western blotting analysis 48 hours after transfection, respectively; the proliferative curves of transfected cells were plotted using MTT assay; furthermore, the change in cellular migration ability was determined using wound healing assay. Results: The eukaryotic expression vector of shRNA targeting S100A6 was successfully constructed. Real-time PCR and Western blotting analysis results showed that S100A6 was effectively silenced by liposome-mediated transfection of the recombinant shRNA vectors in Eca109 cells. Compared with the untransfected cells, S100A6 mRNA and protein in transfected Eca109 cells were significantly decreased (P<0.05, P<0.01). Meanwhile, the proliferative activity of Eca109 cells was significantly inhibited by S100A6 silencing (P<0.01). It was found that the cellular migration was also suppressed by S100A6 gene interference. Conclusion: S100A6 gene can be effectively silenced by shRNA expression vectors, and the silence may lead to inhibition of the proliferation and migration of Eca109 cells.