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Research ArticleExperimental Studies

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

MADOKA NAEMURA, CHIHIRO MURASAKI, YASUTOSHI INOUE, HARUNA OKAMOTO and YOJIRO KOTAKE
Anticancer Research October 2015, 35 (10) 5377-5382;
MADOKA NAEMURA
Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and Engineering, Kinki University, Fukuoka, Japan
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CHIHIRO MURASAKI
Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and Engineering, Kinki University, Fukuoka, Japan
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YASUTOSHI INOUE
Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and Engineering, Kinki University, Fukuoka, Japan
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HARUNA OKAMOTO
Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and Engineering, Kinki University, Fukuoka, Japan
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YOJIRO KOTAKE
Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and Engineering, Kinki University, Fukuoka, Japan
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  • For correspondence: ykotake@fuk.kindai.ac.jp
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    Figure 1.

    Expression levels of ANRIL (antisense non-coding RNA in the INK4 locus) in various human cells. The levels of ANRIL expression were determined by RT-PCR. RPL32 (ribosomal protein L32) was used as an internal control.

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    Figure 2.

    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).

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    Figure 3.

    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).

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    Figure 4.

    Silencing of ANRIL (antisense non-coding RNA in the INK4 locus) causes cell-cycle arrest at the G2/M phase. A: HeLa cells transfected with control (Ctr-i) or ANRIL siRNA oligonucleotides were analyzed by Muse™ Cell Analyzer at 72 h after transfection. B: The percentage of total cells present in the G0/G1, S and G2/M phases of the cell cycle are shown.

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Anticancer Research: 35 (10)
Anticancer Research
Vol. 35, Issue 10
October 2015
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Long Noncoding RNA ANRIL Regulates Proliferation of Non-small Cell Lung Cancer and Cervical Cancer Cells
MADOKA NAEMURA, CHIHIRO MURASAKI, YASUTOSHI INOUE, HARUNA OKAMOTO, YOJIRO KOTAKE
Anticancer Research Oct 2015, 35 (10) 5377-5382;

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Long Noncoding RNA ANRIL Regulates Proliferation of Non-small Cell Lung Cancer and Cervical Cancer Cells
MADOKA NAEMURA, CHIHIRO MURASAKI, YASUTOSHI INOUE, HARUNA OKAMOTO, YOJIRO KOTAKE
Anticancer Research Oct 2015, 35 (10) 5377-5382;
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Keywords

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