Effects of KISS1 gene transfected by lentivirus on proliferation, invasion, and migration abilities of human colorectal cancer HCT116 cells / 吉林大学学报(医学版)

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.

Suppression of Kiss-1 gene inhibits HCT116 human colorectal carcinoma cell migration in vitro via nuclear factor-κB signaling pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway.</p><p><b>METHODS</b>A recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection.</p><p><b>RESULTS</b>In cells over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell proliferation, invasion, and migration as compared to the control cells (P<0.05).</p><p><b>CONCLUSION</b>Lentivirus-mediated Kiss-1 gene over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.</p>

Effect of Kiss-1 gene promoter methylation on its expression in colorectal carcinoma tissue and its clinical significance / 吉林大学学报(医学版)

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.

Association between small heterodimer partner gene and high birth weight infants / 中华围产医学杂志

Objective To evaluate the relationship of small heterodimer partner(SHP) gene and birth weight in China. Methods A cohort study of 191 normal pregnant women was conducted. Both maternal and cord blood samples were collected. PCR-RFLP was used to detect the polymorphism of SNP-rs7504 of SHP. Results (1) The frequency of both neonatal and maternal C allele and (TC+CC) genotype increased significantly with birth weight (P=0.004, OR=3.168; P=0.005, OR=3.315; P=0.013, OR=2.495; P=0.013, OR=2.495). (2) The babies were heavier if they were C allele carrier. The average increase of birth weight was 246.3 g comparing the neonates with TC+CC genotype with those with TT genotype [(3658.7?400.94)g vs (3412.4?444.4)g, P=0.005]. The average birth weight of those maternal C allele carriers was 210.3 g heavier than those non-C allele carriers[(3628.9?405.5) g vs (3418.6?449.0 g]. (3) The fetal C allele was associated with maternal weight in pregnancy, prepregnant BMI, paternal height and weight. Women with C allele were heavier and had higher BMI without statistical significance comparing with those non-C allele carriers. Neither neonatal nor maternal SHP gene was associated with blood glucose and insulin level. (4) Multiple factors analysis showed that birth weight was related to maternal height, weight gain during pregnancy, prepregnant BMI, maternal and cord blood insulin level. After adjustment, the neonatal birth weight remained significantly correlated with cord blood SHP (P=0.0354), but not with maternal SHP gene (P=0.0711). Conclusions SHP gene is associated with newborns birth weight and may affect fetal growth.