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The Effects of LRP16 Gene on the Function of MIN6 Cell Line

Author: XingGuang
Tutor: MuYiMing；WangBaoAn
School: PLA Postgraduate Medical School
Course: Within the scientific endocrine and metabolic diseases, professional
Keywords: LRP16 MIN6 Insulin The glucose transporters sub -2 PDX -1
CLC: R346
Type: Master's thesis
Year: 2009
Downloads: 59
Quote: 0
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Abstract

LRP16 gene is the People's Liberation Army General Hospital, Korea for the East, etc. In 1999, the first cloned a human gene. Is located on chromosome 11q12.23, the expression product is a nucleoprotein. Target genes that previous studies have found that LRP16 gene for estrogen, estrogen through the estrogen receptor α (ERα) increases the gene, while LRP16 while ERα coactivator, feedback enhanced ERα-mediated transcriptional activity. In recent years a large number of studies found that estrogen protect islet β-cells, promote the role of insulin synthesis and secretion, the role through the LRP16 the mediated whether concern. Recent study found that human pancreatic β cell tumors Immunohistochemistry showed that LRP16 protein expression was significantly higher than the normal controls. This suggests that the the LRP16 gene may play an important role in growth, development and islet β-cells to synthesize insulin secretion in the pancreas. This study LRP16 gene overexpression in pancreatic β cell line MIN6 cells, expression of detecting glucose-stimulated insulin secretion function (GSIS) and insulin mRNAs, and to explore its possible mechanism. The study is divided into two parts: the LRP16 gene MIN6 cell proliferation and insulin secretion function Objective: To investigate the proliferation and insulin secretion in of LRP16 gene MIN6 cells function and its possible mechanism. Method: 1. MIN6 cells detected by Western blot whether the expression of LRP16 protein; SuperFect the liposome transfection method to establish MIN6 cell lines stably overexpressing LRP16 gene; 3. Proliferation using the MTT assay; 4. 0 mmol / L, respectively, 3 mmol / L and 30 mmol / L glucose-stimulated cells detect insulin secretory function; 5. using Western blot was used to detect the glucose transporter -2 (Glut-2) protein expression. Results: 1.MIN6 cells express the LRP16 proteins; 2 over-expression the LRP16 group of cell proliferation control group showed no significant difference (p> 0.05); 3. 0 mmol / L, 3 mmol / L and 30 mmol / L glucose-stimulated, over-expression of the LRP16 insulin secretion were 2.26 times that of the control group (p <0.05), 2.19-fold (p <0.05) and 2.16-fold (p <0.05) (among compared, p > 0.05); 4.Westernblot expression the LRP16 group Glut-2 protein amount is 1.76 times of the control group (p <0.05). Conclusion: The expression of LRP16 protein in mouse pancreatic β cells; over-expression of the LRP16 gene can not promote MIN6 cell proliferation, but can promote glucose-stimulated insulin secretion (GSIS), the effects may depend on the upregulation of Glut-2, does not depend on glucose concentration. LRP16 gene impact on the the MIN6 cells insulin mRNAs synthesis and transcription factor Objective: To investigate the the the LRP16 gene synthesis of insulin mRNAs MIN6 cells and may be related transcription factor. Method: 1. With superFect liposome transfection method to establish stable expression of the the LRP16 gene of MIN6 cell lines; 2 Real-Time PCR assay synthetic insulin mRNAs and transcription factor PDX -1 (Pdx-1), MafA and NeuroD1 mRNAs of the synthesis; 3. Pdx-1, MafA and NeuroD1 protein expression was detected by Western blot. Results: 1.Real-Time PCR expressing LRP16 group of Insulin Ⅰ mRNA and Insulin Ⅱ mRNA in the amount of, respectively, is the control group 1.6-fold and 1.8-fold (p-<0.05); 2. Excessive expression of LRP16 group of Pdx-1 mRNA in the amount is in control 1.62 times the group (p <0.05), while MafA and NeuroD1 mRNAs amount compared to the control group, there was no significant difference (p> 0.05); 3 overexpression The LRP16 group Pdx-1 protein amount is 2.04 times that of the control group ( p <0.05), while MafA and NeuroD1 protein amount compared to the control group, there was no significant difference (p> 0.05). Conclusion: Overexpression of the LRP16 gene can promote the synthesis of insulin mRNAs by upregulating transcription factor Pdx-1 implementation.

The Effects of LRP16 Gene on the Function of MIN6 Cell Line

Abstract

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