Figure 5.

HNF1A-AS1 suppresses tumor growth and lymph node metastasis in vivo. (A) Typical specimens in the subcutaneous xenotransplant tumor model of human laryngeal cancer in nude mice. (B) Tumor growth curve and (C) RT-qPCR analysis of HNF1A-AS1 expression in vivo after the TU-686/shRNA-2048, TU-177/HNF1A-AS1 and control cells were injected subcutaneously into the right flank of nude mice (n=20). Tumor growth was calculated from day 5 and tumors were harvested from mice at 43 days after injection. (D) Typical specimens of tumors injected in tongues submucosally in the cervical lymph node metastasis model in nude mice. (E) Total weight of tumors submucosally injected into tongue of nude mice (n=24). (F) RT-qPCR analysis of HNF1A-AS1 expression. Tumors and cervical lymph nodes were harvested from mice at 15 days after injection. (G) Representative HE staining of cervical lymph node metastasis in HNF1A-AS1 knockdown or overexpression cells. Scale bar, 50 µm. Arrows indicate the metastasis in cervical lymph nodes. (H) Ratios of cervical lymph node metastasis in nude mice inoculated submucosally with the indicated cells into the tongue. Data are presented as the mean ± SD. All assays were performed in triplicate, and the values represent the mean of three independent experiments. For statistical analysis, an unpaired t-test was used. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. The xenograft tumorigenicity experiment in vivo was performed one time. HNF1A-AS1, hepatic nuclear factor 1 α antisense RNA 1; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; shRNA, short hairpin RNA.