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Objective:To explore the effects of KISS1 gene transfected by lentivirus on the proliferation,invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms.Methods:The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected.The lentiviral vectors were builted and transfected the KISS1 gene,and the cells were divided into control group (treated with PBS),empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells;Transwell chambers method was used to detect the invasion and migration abilities of the cells.Results:Among LoVo,SW620,SW480,HCT-116,and HT29 cells,the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells,so they were chosen as the research carrier.After transfected with lentiviral vectors,the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene,and the MOI was over 80%.Compared with control group and empty vector group,the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P0.05).Conclusion:The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation,invasion and migration abilities of the colorectal cancer cells,and the mechanism may be related to the expression of KISS1 protein.
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Objective To research the effect of the Kiss-1 gene promoter methylation on the Kiss-1 gene expression in colorectal carcinoma tissue,and to analyze the relationship between the Kiss-1 gene methylation and the clinical pathological features of colorectal carcinoma and its clinical significance.Methods The Kiss-1 gene promotor region methylation,Kiss-1 gene mRNA and protein expressions were detected respectively by methylation-specific PCR, Real-time PCR and Western blotting method in 126 cases of colorectal carcinoma tissue and para-cacinoma normal colorectal tissue.Results The positive rate of Kiss-1 gene methylation in colorectal carcinoma tissue (83.33%)was significantly higher than that in normal tissue (30.16%)(P0.05 ). Conclusion The Kiss-1 gene promoter methylation in colorectal carcinoma tissue is associated with the Kiss-1 gene expression level and the malignant characteristics of colorectal carcinoma;Kiss-1 gene promoter methylation may be used as a reference indicator for predicting the risk of metastasis of colorectal carcinoma.
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Objective To study the diagnosis and treatment of gastric stromal tumor(GST). Methods The clinical manifestation,pathological features ,diagnosis and treatment of 14 patients with GST were retrospectively analysed . Results Of the 14 GST,7 located in the fundus and the body of the stomach, 7 in the antrum of the stomach. The main symptom was abdominal pain, All the 14 GST were diagnosed by endoscopey, barium, ultrasound and CT. All patients were treated by excision. Histological diagnosis was as benign tumor in 8(57.1%)cases, uncertain types in 5 (35.7%)cases , and malignant tumor in 1 ( 7.1 %)case. There were local recurrence in 3 cases and death in 1 case after the operation. Conclusions The complete local excision is recommended for GST patients. Long term postoperative follow up is necessary for patients with GST. Reexcision may be helpful to the patients with recurrence or metastasis.
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Objective To observe the effect of STK15 gene silencing on the growth of gastric cancer cell line-MKN45 in vitro and vivo. Methods STK15 expression was inhibited by RNAi techenique, STK15 protein level was detected by Western blot, the ability of MKN45 invasion in vitro was assessed by cell migration and invasion assay, the change of cell cycle distribution was detected by flowcytometry, MKN45 proliferation was measured by MTT method, and MKN45 cells treated with STK15 siRNA were transplanted subcutanuously in nude mice and their tumorgenesis ability were observed. Results After treatment with STK15 siRNA, STK15 protein level decreased obviously. Compared with control group, STK15- group showed lower invasion ability in vitro [ mean A value: (182?27 ) vs. (308?38 ), P