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Open access

Lin Fang, Hai-Bing Zhang, Hua Li, Yong Fu and Guang-Shun Yang

miR-548c-5p inhibits proliferation and migration and promotes apoptosis in CD90+ HepG2 cells

Background. Since the introduction of the theory of tumour stem cells (TSCs), the liver cancer stem cell (LCSC)-like cells have become one of the focuses in the research on liver cancer.

Materials and methods. In this study, CD90+ cells were applied as the possible LCSC-like cells, and the miRNA and gene expression were analyzed in the CD90+ HepG2 cells. The pilot study showed miR-548c-5p exerted potential effect on the CD90+ HepG2 cells and was thereafter applied for the further study. CD90+ HepG2 cells were assigned to miR-548c-5p mimic transfection group and control group. MTT assay was performed to detect the proliferation of CD90+ HepG2 cells. The migration and invasion abilities were examined by wound healing assay and transwell migration assay, respectively. A detection of apoptosis was performed by fluorescence microscopy.

Results. Our results showed that caspase-3 and bcl-2 were down-regulated while caspase-8 was up-regulated in the CD90+ HepG2 cells. Moreover, the miR-548c-5p transfection could down-regulate the expression of β-catenin, Tg737, bcl-2, bcl-XL, and caspase-3, inhibit the proliferation, migration and invasion and promote the apoptosis of the CD90+ HepG2 cells.

Conclusions. Our findings indicate the imbalance between apoptosis and anti-apoptosis in the LCSC-like cells, which influence the biological features of LCSC-like cells. miRNA plays a regulatory role in the LCSC-like cells among which miR-548c-5p might be a suppressor.

Open access

Ya-Li Liu, Yao-Zhong Ding, Jun-Fei Dai, Bing Ma, Ji-Jun He, Wei-Min Ma, Jian-Liang Lv, Xiao-Yuan Ma, Yun-Wen Ou, Jun Wang, Yong-Sheng Liu, Hui-Yun Chang, Yong-Lu Wang, Qiang Zhang, Xiang-Tao Liu, Yong-Guang Zhang and Jie Zhang

Abstract

Introduction

The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information.

Material and Methods

A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay.

Results

The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results.

Conclusions

A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.