The Expression And Role Of Leptin And Leptin Receptor In Human Papillary Throid Carcinoma Cell Line K1

To study the effect of different concentrations of leptin on the proliferation, migration and invasion on human papillary thyroid carcinoma cell K1. To examine the effect of leptin receptor silencing on the proliferation, migration and invasion on human papillary thyroid carcinoma cell K1.Method Different concentrations of leptin(0 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 125 ng/ml) were used to treat papillary thyroid carcinoma cells K1. The MTT assay, scratch assay and transwell were used to assess the viability and invasive ability of K1 cells respectively. By optimizing transfection conditions that we experimentize and used negative FAM labeled control si RNA. According to the best transfection conditions, the Ob R si RNA vector was transfected into K1 cells by liposome. Three groups including control group, negative control group, Ob R si RNA group were used. K1 cells cultured for 48 hours and 24 hours in 6 well plate and then transiently transfected with si RNAs. After transfection, we detect the expression of target genes at m RNA level and protein level with Real-time PCR and Western Blotting, respectively. MTT was used to detect proliferation of K1 cells after Ob R gene knockout. Transwell was used to detect invasion of K1 cells after Ob R gene knockout.Result 1 The MTT assay showed after K1 cells were treated with different concentrations of leptin, leptin had no effect on the proliferation of K1 cells(P>0.05). After 24 hours the scratch test revealed the migratory rates were: 47.3832%, 59.9498%, 68.7323%, 79.3063%. 48 hours later the migratory rates were: 49.3978%, 74.192%, 86.7326%, 87.7952%, 90.0051%.Transwell assay showed that the number of cells pass through the membrane have an increasing trend(P<0.05). 2 After transient transfection, Real-time PCR showed that the level of m RNA expression decreased on K1 cells.Western Blotting results show that the level of protein expression has targeted inhibition on K1 cells after Ob R gene silencing. MTT results show that the silencing of Ob R has no significant on the proliferation of K1 cells. Transwell results show that the silencing of Ob R decreased K1 cells invasion and the number of cells across the membrane is decreased.Conclusion Leptin and its receptor do not affect human papillary thyroid carcinoma cell K1 proliferation. Leptin enhanced the migratory and invasion in a dose-dependent and time-dependent manner. Ob R si RNA effectively inhibit the expression of leptin receptor on human papillary thyroid carcinoma cells K1 and the ability of invasiveness of K1 cells.