Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or VSports app下载. mil. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

. 2014 Feb;7(2):461-465.
doi: 10.3892/ol.2013.1725. Epub 2013 Dec 3.

miR-101 sensitizes A549 NSCLC cell line to CDDP by activating caspase 3-dependent apoptosis

Jiqing Yin  1 , Mingguo Wang  2 , Cuixiang Jin  1 , Qingguo Qi  3
Affiliations

V体育安卓版 - miR-101 sensitizes A549 NSCLC cell line to CDDP by activating caspase 3-dependent apoptosis

Jiqing Yin et al. Oncol Lett. 2014 Feb.

Abstract

MicroRNA-101 (miR-101) is evidently downregulated in several types of cancer, including non-small cell lung cancer (NSCLC), and is crucial in sensitizing cells to chemotherapy drugs. The aim of the present study was to investigate the correlation between miR-101 and chemosensitivity in the A549 NSCLC cell line. Here, we used the human A549 cell line for transfection with an miR-101 overexpressing vector and detected the cytotoxic acticity, proliferation and apoptosis of cis-diaminedichloroplatinum (CDDP) in A549-miR-101 and A549-mock cells. We demonstrated that overexpression of miR-101 sensitized A549 cells to CDDP, one of the most frequently used agents in curing or controlling NSCLC and enhanced CDDP-induced cell death and caspase 3-dependent apoptosis. In addition, miR-101 facilitated the inhibitory role of CDDP in A549 cell colony formation. Overall, the results of the present study demonstrated that miR-101 sensitizes the A549 NSCLC cell line to CDDP via the activation of caspase 3-dependent apoptosis. VSports手机版.

Keywords: A549; CDDP; NSCLC; chemosensitivity; miR-101; miRNA. V体育安卓版.

PubMed Disclaimer

"V体育ios版" Figures

Figure 1
Figure 1
Effect of the overexpression of miR-101 on the cytotoxic activity of CDDP in A549 cells. (A) Verification of miR-101 transfection. The expression levels of miR-101 were confirmed by real-time polymerase chain reaction. miR-101-transfected cells showed higher miR-101 expression than the Con (P<0.01) and miR-Con (P<0.01) groups. (B) Cell viability was detected by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide assay in pre-miR-101 transfected cells and Con and miR-Con groups following treatment with CDDP for 24 h. Data are presented as the mean ± SEM and experiments were performed in triplicate. *P<0.05 and **P<0.01, between CDDP + miR-101 and CDDP + miR-Con groups. ***P<0.001. miR-101, microRNA-101; Con, untransfected negative control; miR-Con, empty vector-transfected; CDDP, cis-diaminedichloroplatinum.
Figure 2
Figure 2
Overexpression of miR-101 in A549 cells increases CDDP-induced cell death. CDDP-induced cell death was assessed by flow cytometry. Following treatment with CDDP for 24 h, miR-101-transfected cells showed a higher ratio of cell death (93.0%), whereas negative controls exhibited a lower cell death ratio: Con (1.3%), CDDP (39.6%) and CDDP plus miR-Con (41.4%). Con, untransfected negative control; miR-Con, empty vector transfected; CDDP, cis-diaminedichloroplatinum.
Figure 3
Figure 3
Apoptotic analysis. (A) Apoptosis of A549 cells from various treated groups was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Results showed increased positive signals in A549 cells overexpressing miR-101 compared with those in the vector control cells (P<0.01). Data shown are representative of three independent experiments. (B) Number of apoptotic cells, indicated by the bright green fluorescence, and total cell number were quantified using the ImageJ programe, and the rate of apoptosis was subsequently calculated. The results are presented as means ± SD. (C) Western blot analysis results showed increased cleaved caspase 3 expression in A549 cells overexpressing miR-101 compared with that in the vector control cells. ***P<0.001, vs. control and CDDP groups. CDDP, cis-diaminedichloroplatinum; Con, untransfected negative control without CDDP; Non, untransfected control with CDDP; miR-Con, empty vector-transfected.
Figure 4
Figure 4
Overexpression of miR-101 inhibits A549 cell colony formation. Colonies were allowed to grow for 14–21 days and positive colony formation (>50 cells/colony) was counted. The results indicated that overexpression of miR-101 significantly diminished the colony formation rates of A549 cells compared with the mock group. Data are presented as the mean ±SD and experiments were performed in triplicate. **P<0.01 and ***P<0.001. miR-101, microRNA-101; Con, untransfected negative control; CDDP, cis-diaminedichloroplatinum.
See this image and copyright information in PMC

"VSports" References

    1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90. - PubMed
    1. Siegel R, Ward E, Brawley O, Jemal A. Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin. 2011;61:212–236. - PubMed
    1. Kelly K, Crowley J, Bunn PA, Jr, et al. Randomized phase III trial of paclitaxel plus carboplatin versus vinorelbine plus cisplatin in the treatment of patients with advanced non-small-cell lung cancer: a Southwest Oncology Group trial. J Clin Oncol. 2001;19:3210–3218. - PubMed (V体育安卓版)
    1. Lee MW, Kim DS, Min NY, Kim HT. Akt1 inhibition by RNA interference sensitizes human non-small cell lung cancer cells to cisplatin. Int J Cancer. 2008;122:2380–2384. - PubMed (VSports手机版)
    1. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–297. - "V体育2025版" PubMed