Objective:To investigate the inhibitory effect of adenovirus-mediated recombinant Buthus martensii Karsch chloride toxin artifact (Ad-rBmK CTa) on human glioma U251 cells and its related mechanisms.Methods:Groups of 3 titer gradients of 3.5×10 9, 7.0×10 9 and 3.5×10 10 pfu/ml Ad-rBmK CTa were set up and applied to U251 cells for 24, 48 and 72 h, and a blank control group (no cells and Ad-rBmK CTa were added) and a negative control group (only U251 cells were added) were set up at the same time. The virus infection status was observed by laser confocal fluorescence microscopy. The cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle and apoptosis in each group were detected by flow cytometry. The expressions of apoptosis-related proteins bax, bcl-2 and caspase-3 were detected by Western blot. Results:The infection rate of Ad-rBmK CTa was over 90% after acting on U251 cells for 24 h. As the titer of Ad-rBmK CTa increased, the proliferation inhibition rate of U251 cells treated for the same hours gradually increased (all P <0.01); with the extension of time, the proliferation inhibition rate of U251 cells treated with the same titer of Ad-rBmK CTa also gradually increased (all P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, the proportion of cells in G 0/G 1 phase was (40.7±0.8)%, and cells in S phase and G 2 phase accounted for (35.7±0.6)% and (23.6±1.4)%, and the difference was statistically significant ( F = 225.119, P < 0.01). When 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 24, 48 and 72 h, the apoptosis rates were (7.4±1.4)%, (19.2±1.7)% and (22.3±1.7)% ( F = 49.470, P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, compared with the negative control group, the expressions of bax and caspase-3 proteins increased, and the expressions of bcl-2 decreased. Conclusions:Ad-rBmK CTa may act on the DNA damage-induced G 1/S detection site to arrest the cell cycle in G 0/G 1 phase, thus inhibiting the proliferation of U251 cells in vitro. However, its induction of apoptosis in U251 cells is not obvious. The mechanism may be related to the direct or indirect inhibition of chloride ion channels.

Objective:To detect the expression level of long non-coding RNA (lncRNA) nuclear receptor subfamily 2 group F member 2-antisense RNA 1 (NR2F2-AS1) and microRNA-129-5p (miR-129-5p) in glioma tissues and to explore its clinical significance.Methods:The glioma tissues of 103 patients with glioma who underwent surgical treatment in the neurosurgery department of Shanxi Provincial People′s Hospital from January 2015 to September 2016 and 50 normal brain tissues removed due to craniocerebral surgery in the same period were selected as the research objects. Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of lncRNA NR2F2-AS1 and miR-129-5p in tissue samples. The correlation between the two indexes and the relationship between the expression changes of the two and clinicopathological parameters of glioma patients were analyzed. Kaplan Meier curve was used to analyze the 5-year cumulative survival rate of glioma patients with different expression levels of lncRNA NR2F2-AS1 and miR-129-5p. Cox regression analysis was used to analyze the factors affecting the poor prognosis of glioma patients.Results:Compared with normal brain tissue, the relative expression of lncRNA NR2F2-AS1 in glioma was higher and the relative expression of miR-129-5p was lower (all P<0.05). There was significant negative relationship between the relative expression of lncRNA NR2F2-AS1 and miR-129-5p in glioma ( r=-0.756, P<0.05). The expression of lncRNA NR2F2-AS1 and miR-129-5p in glioma was related to World Health Organization (WHO) grade and tumor length (all P<0.05). The 5-year cumulative survival rate of patients in lncRNA NR2F2-AS1<2.89 group was higher than that in lncRNA NR2F2-AS1≥2.89 group, and the 5-year cumulative survival rate of patients in miR-129-5p<0.55 group was lower than that in miR-129-5p≥0.55 group (all P<0.05). Multivariate analysis showed that high expression of lncRNA NR2F2-AS1, low expression of miR-129-5p, WHO grade Ⅲ-Ⅳ and tumor length were the risk factors affecting the prognosis of patients with glioma (all P<0.05). Conclusions:The increased expression of lncRNA NR2F2-AS1 and the decreased expression of miR-129-5p in glioma tissues are involved in the clinical biological progress of glioma and are closely related to the prognosis of patients.