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. 2019 Dec 20;12:11325–11338. doi: 10.2147/OTT.S231846

Figure 6.

Figure 6

HOTTIP regulated HMGA1 through functioning as a ceRNA of miR-218 in GC cells. (A and B) The qRT-PCR analysis was performed to evaluate the subcellular location of HOTTIP in GC cells. (C) Potential binding sites of miR-218 within HOTTIP or HMGA1 were predicted using the miRcode or starBase, respectively. (D) The qRT-PCR assay was carried out to analyze the abundance of miR-218 in cisplatin-resistant and -sensitive GC cells. (E and F) Dual-luciferase reporter assay was conducted to determine the luciferase activity in MGC-803 and MKN-45 cells co-transfected with miR-NC or miR-218 and WT-HOTTIP or MUT-HOTTIP. (G and H) The luciferase activity was measured in MGC-803 and MKN-45 cells co-transfected with miR-NC or miR-218 and HMGA1 3ʹUTR-WT or HMGA1 3ʹUTR-MUT. (I and J) The enrichment of HOTTIP, miR-218, and HMGA1 was measured by RIP assay in MGC-803 and MKN-45 cells incubated with Ago2 or IgG. (K and L) The protein level of HMGA1 was detected in MGC-803 and MKN-45 cells transfected with miR-NC, miR-218, miR-218+ pcDNA, or miR-218+ HOTTIP using the Western blot analysis. *P<0.05.