Dataset Information

RNAi targeting of hTERT gene expression induces apoptosis and inhibits the proliferation of lung cancer cells.

ABSTRACT: The present study aimed to investigate the effects of RNAi-mediated reduction in human telomerase reverse transcriptase (hTERT) expression on apoptosis and lung cancer cell proliferation. A number of cell lines, including 95D, were used. hTERT mRNA levels were detected, and the RNA concentration was calculated. MTT assay was used to detect the inhibition of cell proliferation. The siRNA with the highest suppression rate, siRNA-1, was transfected into 95D cells at three different concentrations (50, 80 and 100 nmol/l). The levels of hTERT mRNA in cells transfected with 50 nmol/l siRNA-1 were not significantly different from those of the negative control-transfected cells (P>0.05), whereas both 80 and 100 nmol/l siRNA-1 showed significant reductions in hTERT mRNA compared to the negative control cells (P<0.01). hTERT levels in the 80- and 100-nmol/l groups were not significantly different (P>0.05). Compared with the control cells, cells transfected with 50, 80 or 100 nmol/l siRNA-1 showed higher fractions of apoptotic cells 48 h post-transfection (P<0.01), although the apoptotic fraction in cells transfected with 50 nmol/l siRNA-1 was not significantly different compared to that in cells transfected with negative control siRNAs (P>0.05). Moreover, the 80- and 100-nmol/l-transfected cells showed significantly increased apoptotic indices (P<0.01). MTT results indicated a time-dependent inhibition of siRNA-1- transfected cell proliferation starting at 12 h and lasting through 48 h post-transfection; the inhibition was attenuated by 72 h post-transfection. The high levels of hTERT mRNA in all human lung cancer cell lines tested suggest that telomerase plays a role in lung carcinogenesis, and this hypothesis was strengthened by the data showing that the siRNA-mediated reduction in hTERT mRNA caused apoptosis and an inhibition of the proliferation of lung cancer cells.

SUBMITTER: Ge L

PROVIDER: S-EPMC3406510 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

RNAi targeting of hTERT gene expression induces apoptosis and inhibits the proliferation of lung cancer cells.

Ge Linhu L Shao Wenlong W Zhang Yangde Y Qiu Yuan Y Cui Dong D Huang Donghai D Deng Zhansheng Z

Oncology letters 20110819 6

The present study aimed to investigate the effects of RNAi-mediated reduction in human telomerase reverse transcriptase (hTERT) expression on apoptosis and lung cancer cell proliferation. A number of cell lines, including 95D, were used. hTERT mRNA levels were detected, and the RNA concentration was calculated. MTT assay was used to detect the inhibition of cell proliferation. The siRNA with the highest suppression rate, siRNA-1, was transfected into 95D cells at three different concentrations ( ...[more]

PMID: 22848277

Similar Datasets

Project description:MicroRNAs (miRNAs) control cell cycle progression by targeting the transcripts encoding for cyclins, CDKs and CDK inhibitors, such as p27(KIP1) (p27). p27 expression is controlled by multiple transcriptional and posttranscriptional mechanisms, including translational inhibition by miR-221/222 and posttranslational regulation by the SCF(SKP2) complex. The oncosuppressor activity of miR-340 has been recently characterized in breast, colorectal and osteosarcoma tumor cells. However, the mechanisms underlying miR-340-induced cell growth arrest have not been elucidated. Here, we describe miR-340 as a novel tumor suppressor in non-small cell lung cancer (NSCLC). Starting from the observation that the growth-inhibitory and proapoptotic effects of miR-340 correlate with the accumulation of p27 in lung adenocarcinoma and glioblastoma cells, we have analyzed the functional relationship between miR-340 and p27 expression. miR-340 targets three key negative regulators of p27. The miR-340-mediated inhibition of both Pumilio family RNA-binding proteins (PUM1 and PUM2), required for the miR-221/222 interaction with the p27 3'-UTR, antagonizes the miRNA-dependent downregulation of p27. At the same time, miR-340 induces the stabilization of p27 by targeting SKP2, the key posttranslational regulator of p27. Therefore, miR-340 controls p27 at both translational and posttranslational levels. Accordingly, the inhibition of either PUM1 or SKP2 partially recapitulates the miR-340 effect on cell proliferation and apoptosis. In addition to the effect on tumor cell proliferation, miR-340 also inhibits intercellular adhesion and motility in lung cancer cells. These changes correlate with the miR-340-mediated inhibition of previously validated (MET and ROCK1) and potentially novel (RHOA and CDH1) miR-340 target transcripts. Finally, we show that in a small cohort of NSCLC patients (n=23), representative of all four stages of lung cancer, miR-340 expression inversely correlates with clinical staging, thus suggesting that miR-340 downregulation contributes to the disease progression.

Dataset Information

RNAi targeting of hTERT gene expression induces apoptosis and inhibits the proliferation of lung cancer cells.

Publications

RNAi targeting of hTERT gene expression induces apoptosis and inhibits the proliferation of lung cancer cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets