Effect of shRNA-mediated inhibition of Nanog gene expression on the behavior of human gastric cancer cells

نویسندگان

  • WEN JI
  • ZHENG JIANG
چکیده

The aim of the present study was to employ RNA interference (RNAi) technology to construct and select shRNA-Nanog recombinant plasmids for the inhibition of Nanog gene expression and transfer these plasmids into the human gastric cancer cell line, SGC-7901, as well as to detect the expression of Nanog and the effects on the proliferation, migration, invasion, cell cycle and apoptosis of SGC-7901 cells. The pshRNA-Nanog interference plasmids were constructed and used to transfect SGC-7901 cells using lipofectamine. The expression of the Nanog gene was detected by fluorescence microscopy, RT-PCR and western blotting, and the most markedly inhibited group was identified. The SGC-7901 cells were transfected with recombinant shRNA-Nanog plasmids from the most markedly inhibited group using lipofectamine and the effect on proliferation was determined by CCK-8 assay. The migration and invasion of the SGC-7901 cells was determined by Transwell assays, while the cell cycle and apoptosis were analyzed by flow cytometry. The group with the highest inhibition rate was successfully constructed and identified. It was observed that the proliferation, invasion and migration capacity of the cells was reduced, that the cell cycle was arrested at the S phase and that apoptosis was significantly increased. The Nanog gene in gastric cancer cells is closely associated with cell proliferation, the cell cycle, apoptosis and migration and invasion abilities. The present study establishes the foundations for a novel approach for the genetic treatment of gastric cancer.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

STUDY OF HMGA2 GENE INHIBITION WITH SPECIFIC SHRNA AND SIRNA AND INVESTIGATION OF CORRESPONDING EFFECTS ON DOWNSTREAM GENE EXPRESSION IN MDA-MB-231 CANCER CELLS: A BIOINFORMATIC AND EXPERIMENTAL STUDY

Background & Aims: The use of siRNA to silence gene expression is increasingly expanding today. The aim of this study is to bioinformatically and experimentally investigate the inhibition of the HMGA2 gene and its corresponding effects on downstream genes expression rate in MDA-MB-231 cancer cell treated by shRNA and siRNA specific to HMGA2. Materials & Methods: To perform this bioinformatic a...

متن کامل

Effect of EZH2 Inhibition on Colorectal Cancer Cells: an In Vitro Study

Recently, the epigenetic modifications have been recognized as a regulator of gene expression in various cancers. EZH2 gene is one the most important component of the PRC2 complex. Overexpression of EZH2 was identified in multiple cancers that considered more attractive the EZH2 role as an oncogene. Some studies report that EZH2 contributes to various aspects of colorectal cancer (CRC). However...

متن کامل

Suppressive Effect of Constructed shRNAs against Apollon Induces Apoptosis and Growth Inhibition in the HeLa Cell Line

Background: Cervical cancer is the second most common female cancer worldwide. Inhibitors of apoptosis proteins (IAPs) block apoptosis; therefore, therapeutic strategies targeting IAPs have attracted the interest of researchers in recent years. Apollon, a member of IAPs, inhibits apoptosis and cell death. RNA interference is a pathway in which small interfering RNA (siRNA) or shRNA (short hairp...

متن کامل

shRNA-mediated downregulation of α-N-Acetylgalactosaminidase inhibits migration and invasion of cancer cell lines

Objective(s): Extracellular matrix (ECM) is composed of many kinds of glycoproteins containing glycosaminoglycans (GAGs) moiety. The research was conducted based on the N-Acetylgalactosamine (GalNAc) degradation of ECM components by α-N-acetylgalactosaminidase (Nagalase) which facilitates migration and invasion of cancer cells. This study aims to investigate the effects of Naga-shRNA downregula...

متن کامل

ShRNA-mediated knock-down of CD200 using the self-assembled nanoparticle-forming derivative of polyethylenimine

Objective(s): ShRNA-mediated silencing strategy is considered to be a potent therapeutic approach. The present study aimed to assess the ability of the previously prepared polyethylenimine (PEI) derivative for the shRNA knock-down of the CD200 gene on the cells obtained from the patients with chronic lymphocytic leukemia (CLL). Materials and Methods: Since there are several investigations...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 6  شماره 

صفحات  -

تاریخ انتشار 2013