[The Detection of Micro RNA346 Gene Polymorphism by Capillary Electrophoresis].

OBJECTIVE: To develop a method for the detection of micro RNA346 gene polymorphism by capillary electrophoresis (CE). METHODS: The genome DNA was extracted with the kit of blood/cell/tissue genome DNA extraction,then micro RNA346 gene was amplified by PCR,digested by BciT130Ⅰrestriction enzyme and detected by CE. The conditions for CE separation were optimized. Samples from rheumatoid arthritis patients and healthy persons were detected under the optimal conditions. RESULTS: Under the optimized experimental conditions of CE (sieving medium mass concentration was 10 g/L and the separation voltage was 12 kV),the detection of the digested products of microRNA346 gene could be completed within 25 min. The intra-day relative standard deviation (RSD) of the method was 0.43%-0.63% and inter-day RSD was 1.49%-1.56%.Samples from 96 rheumatoid arthritis patients and 43 healthy persons were analyzed by the proposed method. The results showed that only micro RNA346Ⅰtype was detected but micro RNA346 Ⅱ type wasn't. CONCLUSION: This method is easy to operate,and has the advantages of high efficiency,fast speed,less sample consumption and high automation level. This method is suitable for the determination of RNA gene polymorphism of mirco RNA.

Circ­TOP2A acts as a ceRNA for miR­346 and contributes to glioma progression via the modulation of sushi domain­containing 2.

Dysregulated circular RNAs (circRNAs) are involved in the carcinogenesis and progression of multiple human malignancies. Knowledge of circRNAs in glioma (GM) is limited and further study to uncover new therapeutic targets for GM is urgently required. The present study demonstrated that circ­TOP2A was elevated in GM tissue specimens and cells and that circ­TOP2A levels indicated an unfavorable clinical prognosis in GM. Functionally, circ­TOP2A knockdown reduced viability, migration and invasion and triggered apoptosis in LN229 cells. Ectopic expression of circ­TOP2A aggravated these malignant behaviors in U87MG cells. In terms of mechanism, RNA­seq was performed to discover the potential targets regulated by circ­TOP2A. Circ­TOP2A acted as a competing endogenous RNA to upregulate sushi domain­containing 2 (SUSD2) expression by sponging microRNA (miR) 346. Rescue assays revealed that the oncogenic function of circ­TOP2A was partially dependent on its regulation of the miR­346/SUSD2 axis. In conclusion, the present study identified that circ­TOP2A promoted GM proliferation and aggressiveness via miR­346/SUSD2 signaling, which is a potential prognostic biomarker and therapeutic target for GM.