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Expressions of PDCD5 in Human Colon Carcinoma and Its Influence of Drug

Author: LiuYanPing
Tutor: WangShiYong
School: China Medical University
Course: Oncology
Keywords: PDCD5 gene Human colon carcinoma HCT116 Immunohistochemical Lentivirus carrier Transfection genes Apoptosis
CLC: R735.3
Type: Master's thesis
Year: 2010
Downloads: 67
Quote: 0
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Abstract


ObjectivePDCD5 gene is China’s own new features gene. A number of laboratories with different technology found, less express of PDCD5 in many types of cancer such as cervical cancer, liver cancer and multiple myeloma, stomach, lung cancer and other diseases, and may be related of its occurrence. Colorectal cancer is a common one of gastrointestinal cancers, in recent years the incidence of colorectal cancer is increasing trend. Cacinoma decelops with typical multiple-stage, colorectal cancer is the most one. In order to study PDCD5 gene expression in Colorectal and its role in development, we designed this experiment.From the organization level, cell protein levels and mRNA level explicitly PDCD5 gene in patients with colorectal cancer disease expression, first look it in colorectal cancer. While on their clinical outcome do follow-up investigation, clear their clinical significance. PDCD5 in tumor cell apoptosis functional research, the use of stable transfection of lentivirus as a carrier, will Lentivirus-mediated PDCD5 transferred to colon cancer HCT116, and to prove that Lentivirus-mediated PDCD5 gene transfer sensitizes HCT116 cells to apoptosis induced by 5-fluorouracil (5-FU) in vitro.MethodsThis investigation detect PDCD5 express level which is utilized by immunohistochemistry on colon carcinoma tissue from 46 cases、adenomatous polyp tissue from 20 cases and chronic enteritis tissue from 28 cases, ask the present conditions with phone call following-up among the 46 colon carcinoma patients. Detection the expression of PDCD5 gene by immunocytochemistry and RT-PCR method respectively in the cell level and mRNA level form colorectal cancer cell lines and normal cell, HCT116 cells were stabily transfected of L-PDCD5 and L-null as control. Cell proliferation rate and viability were measured by MTT and FCM.Analyze the function of PDCD5 gene in the tumor. ResultPDCD5 expression level is the highest in chronic enteritis tissue, significantly higher than the others (χ2=10.495, P＜0.05),but no significant difference between adenomatous polyps and cancer tissues(P= 0.552); In follow-up cases, patients with high expression of PDCD5, survival rates is higher. The express of PDCD5 in Huvec is higher than that in colon cancer cell lines. L-PDCD5 and L-null were transfected into HCT116. GFP can be observed by Fluorescence microscopy, transfection rate is 80-90%.PDCD5 mRNA expression were obvious upregulated in L-PDCD5 transfected HCT116 cells than control vector transfected cells, which illuminates cell transfection of L-PDCD5 successfully. MTT results show the inhibition rate of L-PDCD5 group is higher than the L-null group and simple cell groups(P<0.05).FCM assay results showed HCT116 cells apoptosis rate of L-PDCD5 transfection was increased significantly compared with L-null transfected HCT116 cells. As the drug concentration increases, the rate of increase apoptosis.ConclusionPDCD5 may be a new management target of colon carcinoma’s early diagnosis and management.Lentivirus-mediated PDCD5 gene could transfer HCT116 cells successful, and the expression of PDCD5 in HCT116 cells is stable.This stable expression sensitizes HCT116 cells to apoptosis induced by chemotherapy drugs with concentration-dependence in vitro.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Intestinal neoplasms
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