Guang-chao Zhang, Yuan Gao, Bing Wang and Yong-hui Lai
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.
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
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.
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.
A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.