ABSTRACT
AIM:To investigate the expression of miR-23a and epithelial splicing regulatory protein 1(ESRP1) in rectal cancer tissues and cell lines as well as their effects on rectal cancer cell viability and apoptosis.METHODS:The relative levels of miR-23a in the rectal cancer tissues and cultured cells were assessed by RT-qPCR.The positive expression of ESRP1 in the rectal cancer tissues and non-cancer tissues was detected by immunohistochemical staining.The sequences of miR-23a inhibitor and inhibitor negative control (NC) were synthesized, and transfected into the SW480 cells.The cell viability was measured by CCK-8 assay.The apoptotic rate was analyzed by flow cytometry.The cell invasion was evaluated by Matrigel counting assay.The expression of ESRP1 was determined by Western blot.The wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ESRP1-3'UTR) plasmid and miR-23a inhibitor or inhibitor NC were co-transfected into the HEK293 and SW480 cells.The dual luciferase activity was detected according to Promega dual luciferase reporter gene assay kit instructions.The cell viability and apoptosis were evaluated by CCK-8 assay and flow cytometry analysis, respectively, after the SW480 cells were transfected with ESRP1 mimic or mimic NC.The expression of ESRP1, caspase-3, Smac and X-linked inhibitor of apoptosis protein (XIAP) in the SW480 cells was detected by Western blot.RESULTS:The expression of miR-23a was significantly up-re-gulated in the rectal cancer tissues and cell lines, while the positive expression of ESRP1 was significantly decreased in the rectal cancer specimens.The miR-23a expression was also closely related to lymphnode metastasis and TNM stages of rectal cancer patients.ESRP1 was inversely correlated with miR-23a in the rectal cancer tissues.After transfection with miR-23a inhibitor in human rectal cancer SW480 cells, the down-regulation of miR-23a induced significant inhibition of cell viability as compared with the cells transfected with inhibitor NC (P<0.01).Furthermore, the apoptotic rate induced by the miR-23a inhibitor transfection was markedly higher than that of control (P<0.01).Luciferase assay showed that ESRP1 was a direct target gene of miR-23a.The cell viability and apoptosis were inhibited and promoted, respectively, after transfection with ESRP1 mimic in the SW480 cells.Promoted expression of ESRP1 significantly up-regulated the levels of caspase-3 and Smac as well as down-regulated the expression of XIAP in the SW480 cells.CONCLUSION:The expression of miR-23a is significantly associated with the growth and apoptosis of human rectal cancer cells by targeting ESRP1.miR-23a may be a potential therapeutic target for the treatment of rectal cancer in the future.
ABSTRACT
Objective To elucidate the relative level of miR-23a RNA in rectal cancer tissues and cell line as well as the effects of miR-23a on the cell proliferation and apoptosis of rectal cancer cells in vitro.Methods Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was applied in assessment of the transcription of miR-23a in rectal cancer tissues and in vitro cells.The RNA fragment of miR-23a inhibitor and inhibitor NC were synthesized and transfected into SW480 cells.Cell proliferation was evaluated with Cell Counting Kit-8 (CCK-8) assay.The apoptotic rate was analyzed by flow cytometry.The expression of ESRP1 was detected by western blot.Wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ES-RP1-3'UTR) plasmids and miR-23a inhibitor RNA fragments or inhibitor NC RNA fragments were co-transfected into HEK293 and SW480 cells,then the Promega dual luciferase reporter gene assay kit was used to examine the dual luciferase activity in SW480 cells.Resuits The relative RNA level of miR-23a was significantly promoted in both rectal cancer tissue samples and SW480 cells.After SW480cells were transfected with miR-23a inhibitor,human rectal cancer cell line SW480 with down-regulation of miR-23a showed significant inhibition of cell proliferation compared with negative control (P =0.000).Furthermore,our data demonstrated clearly that the inhibition of miR-23a promoted apoptosis in SW480 cells (P =0.000).Luciferase assay showed that ESRP1 was a direct target gene of miR -23a.Conclusion The expression of miR-23a is clearly associated with the growth and apoptosis of human rectal cells by targeting ESRP1,whilst miR-23a may be used as a potential therapeutic target for the treatment of rectal cancer in the future.
ABSTRACT
Objective To explore the clinical effects of single-agent Xeloda (Capecitabine) therapy and the related risk factors in patients with advanced colorectal cancer. Method Seventy-eight patients with advanced colorectal cancer were treated with oral Xeloda, 1250 mg/m 2 twice daily, on days 1-14 every 21 days. At least 2 cycles were administered. The short-term clinical effects were evaluated, and the related risk factors were tested by Logistic regression analysis. Results The overall response rate was 32.05%with 5 cases complete response (CR), 20 cases partial response (PR), 31 cases stable disease (SD), 22 cases progress disease (PD). The Logistic regression analysis showed that the age (OR=1.52, 95%CI 1.015~2.319), fast blood glucose (OR=1.30, 95%CI 1.483~3.677), albumin (OR=1.98, 95%CI 1.526~2.572), ALT (OR=2.37, 95%CI 1.621~3.509) and AST (OR=2.21, 95%CI 1.526~2.572) were independent risk factors for inefficient treatment. Conclusion The single-agent Xeloda (Capecitabine) is an efficacious treatment for the patients with advanced colorectal cancer. However, the inefficient rate is also high and it relates to a variety of factors. We should comprehensively evaluate the patients to improve the short-term clinical effects.