Fig. 3.

Knockdown of long noncoding RNA HEIH (lncRNA-HEIH) inhibits colorectal cancer tumorigenesis. (A) The expression of lncRNA-HEIH in lncRNA-HEIH stably knocked-down and control LoVo cells was detected by quantitative real-time polymerase chain reaction and normalized to glyceraldehyde 3-phosphate dehydrogenase. (B) Cell proliferation rate of lncRNA-HEIH stably knocked-down and control LoVo cells were detected by the Cell Counting Kit-8 assays. (C) Proliferative cells of lncRNA-HEIH stably knocked-down and control LoVo were labeled with ethynyl deoxyuridine (EdU). Red color indicts EdU-positive cells. Scale bars=100 μm. (D) The level of apoptosis in lncRNA-HEIH stably knocked-down and control LoVo cells was detected by TdT-mediated dUTP nick end labeling (TUNEL) staining. Blue color indicts TUNEL-positive cells. Scale bars=100 μm. For A-D, results are shown as mean±standard deviation (SD) from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t test. (E, F) lncRNA-HEIH stably knocked-down and control LoVo cells were subcutaneously injected into nude mice. Tumor volumes were detected every 7 days (E). Tumor weights were detected at the 28th day after injection (F). (G) Tumors generated from subcutaneous injection with lncRNA-HEIH stably knocked-down and control LoVo cells were immunohistochemistry stained for Ki-67. Scale bars=50 μm. (H) The same tumors as in panel G were stained for TUNEL. Scale bars=100 μm. For E-H, results are shown as mean±SD from six mice. **p < 0.01 by Mann-Whitney U test.