Let-7a-5p inhibits HK2 expression by directly targeting its 3'-UTR in oral squamous cell carcinoma cells. (IMAGE)
Caption
(A, B) Immunoblot analysis of HK2 expression in CAL27 and SCC9 cells transfected with non-targeting control (NTC), (A) let-7a-5p mimics, or (B) let-7a-5p antagomirs. Histograms show relative let-7a-5p expression by quantitative reverse transcription PCR. (C) HK2 mRNA levels were measured by quantitative reverse transcription PCR in CAL27 and SCC9 cells expressing NTC or let-7a-5p mimics. (D) The top panel indicates wild-type (WT) and mutated (Mut) forms of putative let-7a-5p target sequences of HK2 3'-UTR. The red font indicates the putative let-7a-5p binding sites within human HK2 3'-UTR. The blue font indicates the mutations introduced into the HK2 3'-UTR. Let-7a-5p mimics were co-transfected with pmirGLO-HK2-3'-UTR or pmirGLO-HK2-3'-UTR-mut into CAL27 and SCC9 cells, followed by dual-luciferase analysis. (E) NTC or let-7a-5p antagomirs were transfected into CAL27 and SCC9 cells with HK2 knockdown, followed by measurement of cellular glucose uptake and lactate production in culture media. The data were presented as mean ± standard deviation of three independent experiments by student's t-test. *P < 0.05, **P < 0.01.
Credit
Shanghui Zhou, Jingyu Zhan, Jia Wang, Jingang Yang, Dahe Zhang, Zhenming Li, Yue He
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