ABSTRACT. The aims of this study were to explore the correlation between the expression of EpCAM and the Wnt/β-catenin pathway in human colon cancer and its clinical significance for the evaluation of cancer prognosis. Samples from colon cancer, para-carcinoma, or benign intestinal tissue from individual patients (50) and from normal intestinal mucosal tissues (20) were obtained from the Pathology Department of the Shandong Province Binzhou People's Hospital (Shandong, China). Immunohistochemistry was used to detect the expression levels of EpCAM and β-catenin proteins in these tissues, and the prognoses of the patients from whom the samples were derived were determined on follow-up examination. The corresponding in vitro mechanistic siRNA experiments were subsequently performed in the human colon cancer cell line HCT116 to observe the regulatory effects of silencing EpCAM expression on the Wnt/β-catenin pathway. From these analyses, we determined that the expression levels of EpCAM and β-catenin were higher in cancer tissues compared with other tissues 4486©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (2): 4485-4494 (2015) from the same patient, and that the expression of EpCAM and Wnt/β-catenin in colon cancers were positively correlated. The prognostic analysis showed an inverse correlation between EpCAM and Wnt/β-catenin expression and patient prognosis. A further examination of cellular mechanisms confirmed that the silencing of EpCAM led to decreased expression of Wnt/β-catenin, and thus reduced proliferation and increased the apoptosis ratio in the cells. These results suggest that suppression of EpCAM might be a new approach for treating colon cancer.

Objectives A small subpopulation of colorectal cancer stem cells (CSCs) possess the ability to self-renew and the capacity to initiate the original tumor. EpCAMhigh/CD44+ cells are regarded as CSCs in colorectal cancer. The present study was undertaken to investigate the significance of EpCAM in the in vitro proliferation ability and oxaliplatin chemoresistance of EpCAMhigh/CD44+ colorectal CSCs. Methods We applied fluorescence-activated cell sorting (FACS) to separate the EpCAMhigh/CD44+ subset from human colorectal cancer cell line HCT116. We also used siRNA targeting EpCAM to create EpCAM−/CD44+ subpopulation. Then we compared EpCAMhigh/CD44+ cells and EpCAM−/CD44+ cells for proliferation ability and the chemoresistance to oxaliplatin by CCK8 assay. Results The EpCAMhigh/CD44+ subset comprises almost 6.25 ± 0.09% in cell line HCT116, and the EpCAM−/CD44+ cells displayed a significantly lower proliferation ability and weaker oxaliplatin chemoresistance than the EpCAMhigh/CD44+ cells. Conclusions EpCAM is critical for tumor proliferation and oxaliplatin chemoresistance in EpCAMhigh/CD44+ colorectal CSCs.

Aims: The present study investigated the exact proportion, the extent of in vitro proliferation potential, and oxaliplatin chemoresistance of EpCAMhigh/CD44+ cancer stem cells in colorectal cancer. Its underlying mechanism was also explored. Background: Colorectal cancer stem cells (CSC) play crucial roles in tumorigenicity and chemoresistance. Multiple studies have shown that JAK/STAT, NOTCH, and Wnt/-catenin pathways, associated with tumour recurrence and metastasis, contribute to the proliferation and maintenance of CSCs. CSCs become resistant to chemo-radiotherapies by improving DNA damage repair, changing cell cycle checkpoints, and scavenging reactive oxygen species, resulting in a bad patient prognosis. Objective: This work was carried out to determine the precise fraction, the degree of in vitro proliferation capability, and the level of oxaliplatin chemoresistance exhibited by EpCAMhigh/CD44+ cancer stem cells in colorectal cancer. The research was also done to investigate its underlying process. Methods: Fluorescence-activated cell sorting (FACS) was applied to isolate the EpCAMhigh/CD44+ populations from three human colorectal cancer cell lines (HCT116, HT29, and LoVo), and we quantified the average proportion of the EpCAMhigh/CD44+ cells in every cell lines. The comparison of their proliferation ability and the chemoresistance to oxaliplatin with the parental cells was estimated by CCK8 assay. The activated signaling pathway was tested by Western Blotting. result: EpCAMhigh/CD44+ subpopulation comprise about 4.98±1.24% of the total human colorectal cancer cell lines, and the EpCAMhigh/CD44+ cells exhibited a highly better proliferation ability and stronger oxaliplatin chemoresistance than the parental cells. Wnt/β-catenin signaling pathway is activated in EpCAMhigh/CD44+ HCT116 cells. Results: EpCAMhigh/CD44+ subpopulation comprises about 4.98±1.24% of the total human colorectal cancer cell lines, and the EpCAMhigh/CD44+ cells exhibited a highly better proliferation ability and stronger oxaliplatin chemoresistance than the parental cells. The wnt/β-catenin signaling pathway is activated in EpCAMhigh/CD44+ HCT116 cells conclusion: Activation of Wnt/β-Catenin signaling in EpCAMhigh/CD44+ cells endow colorectal cancer with tumor proliferation and oxaliplatin chemoresistance. Conclusion: Activation of Wnt/β-Catenin signaling in EpCAMhigh/CD44+ cells endow colorectal cancer with tumor proliferation and oxaliplatin chemoresistance.

BackgroundPrior study indicates that abnormal protein expression and functional changes in the development and progression of colorectal cancer is related to gene expression. The aim of this study was to construct an interference plasmid targeting the Ep-CAM gene and to investigate its effects on the proliferation of colorectal cancer cells.MethodsIn this study, HT-29 and HCT-116 colorectal cancer cell lines were selected as cell models. The double-stranded micro (mi)RNA oligo was inserted into the pcDNATM6.2-GW/EmGFPmiR vector, which is an expression of miRNA. Lipofectamine™ 2000 was used to transfer plasmid into the empty plasmid group (transfected pcDNATM6.2-GW/EmGFPmiR-neg) and the interference group (transfected pcDNATM6.2-GW/EmGFPmiR-Ep-CAM-1), respectively. Meanwhile, the nontransferred HT-29 and HCT-116 acts as the blank control group. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the transfection efficiency. Western blot was used to detect Ep-CAM protein expression. The cell proliferation in each group was detected by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.ResultsThe results indicated that the Ep-CAM messenger (m)RNA expression in the interference group was lower significantly compared with that of the empty plasmid group and control group (P<0.01). Western blot analysis results showed that Ep-CAM protein expression was significantly lower in interference group compared with that of the empty plasmid group and the control group (P<0.01). MTT assay results demonstrated that the proliferation ability of cells in the interference group was significantly inhibited compared with the two other groups (P<0.05).ConclusionSilencing of Ep-CAM can significantly inhibit the proliferation of colorectal cancer cells.

Objective: To explore the clinical effect of domestic capsosol foam sclerotherapy in the treatment of hemorrhoids under transparent cap assisted endoscope. Methods: According to the experimental requirements, 120 cases of hemorrhoids hospitalized in gastroenterology department in our hospital from February to December in 2019 were selected as the treatment group. The foam sclerotherapy of polidocanol and the air in accordance with the 1:4 ratio was injected into varicose hemorrhoids and hemorrhoids through a transparent cap assisted endoscope. At the same time, 100 patients were selected as the control group and the patients were treated according to the routine protocol. The symptom score and clinical efficacy of the two groups were observed under different treatment schemes. Results: 120 patients were successfully injected with foam sclerotherapy under endoscope. There were significant differences in clinical symptom score and clinical efficacy between the injection sclerotherapy group and the control group. There were statistically significant differences in the bleeding, prolapse, painful defecation, anal foreign body sensation, impact on daily work, and hemorrhoids data between the treatment group and the control group (P<0.05). Conclusion: Transparent cap assisted endoscopic injection of polidocanol sclerotherapy in the treatment of hemorrhoids can effectively improve the clinical symptoms of patients, promote the improvement of the disease, and the curative effect is satisfactory. It is a new method of minimally invasive treatment of hemorrhoids, which is worthy of wide promotion in clinical practice.