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. 2021 Feb 3;21(4):317. doi: 10.3892/etm.2021.9748

Figure 4.

Figure 4

miR-136-5p overexpression increases the cisplatin sensitivity of laryngeal squamous cell carcinoma and head and neck squamous cell carcinoma cells. (A) After cells were transfected with miR-136-5p mimics or mimics-NC and treated with cisplatin (2.6 µM) for 24 h, cell viability was detected using MTT assays, n=6. (B) Flow cytometric detection of apoptosis using annexin V-FITC/PI staining and (C) quantification of apoptosis. Autophagy was detected using an autophagy double-labeled adenovirus infection assay in (D) FD-LSC-1 and (E) FaDu cells, scale bar=50 µm. Free yellow puncta indicates the autophagosome, free red puncta indicates the autophagolysosome. (F) The protein expression levels of ROCK1, p-Akt/Akt (Ser473), GSK-3β, p-GSK-3β (Ser9), p-mTOR (S2448)/mTOR, LC3II/I and P62 were measured using western blot analysis and normalized to the levels of β-actin. Data are presented as the mean ± SD, n=3. *P<0.05. GSK-3β, glycogen synthase kinase-3β; LC3, microtubule-associated protein 1 light chain 3; miR, microRNA; mTOR, mammalian target of rapamycin; NC, negative control; p-, phosphorylated.