Fig. 2
The effects of MRVI1-AS1 on metastasis and growth of HCC cells. A MRVI1-AS1 shRNAs (shMRVI1-AS1#1, shMRVI1-AS1#2) significantly decreased the expression of MRVI1-AS1 in MHCC-97H cells (mean ± SD; n = 3). ***P < 0.001, two-way ANOVA. The pcDNA/MRVI1-AS1 significantly increased the expression of MRVI1-AS1 in Hep3B cells (mean ± SD; n = 3). *P < 0.05, Student’s t test. B, C Transwell assays revealed that MRVI1-AS1 shRNAs notably suppressed migration and invasion of MHCC-97H cells (mean ± SD; n = 3). ***P<0.001, two-way ANOVA. MRVI1-AS1 overexpression notably promoted migration and invasion of Hep3B cells. (mean ± SD; n = 3). ***P < 0.001, Student’s t test. D, E Wound healing assays revealed that MRVI1-AS1 silencing markedly suppressed MHCC-97H cells mobility (mean ± SD; n = 3). ***P < 0.001, two-way ANOVA. Ectopic expression of MRVI1-AS1 obviously strengthened migration of Hep3B cells (mean ± SD; n = 3). **P < 0.01, Student’s t test. F, G MTT assays indicated that MRVI1-AS1 knockdown notably suppressed viability of MHCC-97H cells. Ectopic expression of MRVI1-AS1 markedly enhanced viability of Hep3B cells (mean ± SD; n = 3). **P<0.01, ***P<0.001, two-way ANOVA. H, I EdU assays revealed that MRVI1-AS1 silencing notably inhibited proliferation of MHCC-97H cells (mean ± SD; n = 3; ***P<0.001, two-way ANOVA), while MRVI1-AS1 overexpression had the contrary effect on Hep3B cells (mean ± SD; n = 3; ***P < 0.05, Student’s t test). J Nude mice were injected by MHCC-97H subclones through the tail vein. Then formation rate of tumor nodule in the lung was evaluated by lung section H&E staining (mean ± SD; n = 5). ***P < 0.001, Student’s t test. K, L MHCC‑97H subclones were injected subcutaneously into the right flank of nude mouse and tumor volume was measured every 7 days (mean ± SD; n = 5). ***P < 0.001, two-way ANOVA with Sidak’s t test. M Tumors were harvested on day 28 and weighed (mean ± SD; n = 5). **P < 0.01, two-way ANOVA. N Tumor RNA was isolated and RT-qPCR analysis of MRVI1-AS1 was performed. ***P < 0.001, Student’s t test. O Tumor sections were analyzed by immunohistochemistry for Ki-67, and the stained area in 10 fields was quantified by Image J software and the percentage of total area that was positive for staining is shown. ***P < 0.001, Student’s t test