Long Noncoding RNA ANRIL Regulates Proliferation of Non-small Cell Lung Cancer and Cervical Cancer Cells

Silencing of ANRIL (antisense non-coding RNA in the INK4 locus) represses the proliferation of HeLa and H1299 cells. A: HeLa cells were transfected with control (Ctr-i) or ANRIL siRNA oligonucleotides. At 72 h after transfection, cells were harvested and subjected to RT-PCR (reverse transcription-polymerase chain reaction) to determine the level of ANRIL. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as an internal control. B: HeLa cells transfected with control or ANRIL siRNA oligonucleotides were observed by phase-contrast microscopy at 72 h after transfection. C: After initial seeding of 5×10⁵ cells, HeLa cells were incubated overnight, and then transfected with control or ANRIL siRNA oligonucleotides. At 72 h after transfection, viable HeLa cells were counted by Trypan Blue staining. D: H1299 cells were transfected with control (Ctr) or ANRIL siRNA oligonucleotides. At 72 h after transfection, cells were harvested and subjected to RT-PCR as in (A). E: H1299 cells transfected with control or ANRIL siRNA oligonucleotides were observed as in (B). F: After initial seeding of 1×10⁵ cells, H1299 cells were incubated overnight, and then transfected with control or ANRIL siRNA oligonucleotides. Viable H1299 cells were counted as in (C).

Silencing of ANRIL (antisense non-coding RNA in the INK4 locus) increases p15 mRNA expression. A: HeLa cells were transfected with control (Ctr-i) or ANRIL siRNA oligonucleotides. At 72 h after transfection, cells were harvested. The effects of ANRIL silencing on the expression of p15, p16 and ARF (alternative reading frame) were determined by qRT-PCR. The results are expressed relative to the corresponding values for control cells. The mean values and standard deviations were calculated from the data of three independent experiments. B: The effects of ANRIL silencing on the expression of p15 and ARF in H1299 cells were determined by qRT-PCR as in (A).