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Published in: BMC Cancer 1/2014

Open Access 01-12-2014 | Research article

microRNA-30b/c inhibits non-small cell lung cancer cell proliferation by targeting Rab18

Authors: Keng Zhong, Kun Chen, Lin Han, Bailing Li

Published in: BMC Cancer | Issue 1/2014

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Abstract

Background

MicroRNAs (miRNAs) are small noncoding RNAs that post-transcriptional regulate gene expression in a variety of cancers. Increasing evidences indicate that miR-30 expression is down-regulated in numerous human cancers including non-small cell lung cancer (NSCLC) which hypothesizes that miR-30 may play an important role in tumorigenesis. The aim of this study was to investigate the target gene of miR-30 and its roles in tumor growth of NSCLC.

Methods

Luciferase reporter assays were employed to validate regulation of a putative target of miR-30. The effect of miR-30 on endogenous levels of this target were subsequently confirmed via Western blot (WB). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to determine the expression level of miR-30 in NSCLC specimens and adjacent non-tumor tissues. MTT assays were conducted to explore the impact of miR-30 overexpression on the proliferation of human NSCLC cells.

Results

Both miR-30b and miR-30c (miR-30b/c) were found having target site in same region of Rab18 mRNA. Luciferase assays using a reporter carrying a putative miR-30b/c target site in the coding DNA sequence (CDS) region of Rab18 revealed that miR-30b/c directly targeted Rab18. Overexpression of miR-30b/c led to down-regulation of Rab18 in A549 and H23 cells at protein levels but not mRNA levels. Down-regulation of miR-30b/c and up-regulation of Rab18 protein levels were detected in NSCLC specimens compared with adjacent non-tumor tissues. Overexpression of miR-30b/c suppressed NSCLC cells growth. Knockdown of Rab18 by siRNA significantly inhibited the proliferation of NSCLC cells.

Conclusions

We demonstrated that miR-30b/c was down-regulated in NSCLC specimens compared with adjacent non-tumor tissues. miR-30b/c directly targeted and down-regulated Rab18 expression and inhibited NSCLC cells proliferation. These data indicated that miR-30b/c could serve as a tumor suppressor gene involved in NSCLC pathogenesis.
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Literature
1.
go back to reference Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61 (2): 69-90. 10.3322/caac.20107.CrossRefPubMed Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61 (2): 69-90. 10.3322/caac.20107.CrossRefPubMed
2.
go back to reference Wood AJJ, Spira A, Ettinger DS: Multidisciplinary management of lung cancer. N Engl J Med. 2004, 350 (4): 379-392. 10.1056/NEJMra035536.CrossRef Wood AJJ, Spira A, Ettinger DS: Multidisciplinary management of lung cancer. N Engl J Med. 2004, 350 (4): 379-392. 10.1056/NEJMra035536.CrossRef
3.
go back to reference Yang L, Parkin DM, Ferlay J, Li L, Chen Y: Estimates of cancer incidence in China for 2000 and projections for 2005. Cancer Epidemiol Biomark Prev. 2005, 14 (1): 243-250. Yang L, Parkin DM, Ferlay J, Li L, Chen Y: Estimates of cancer incidence in China for 2000 and projections for 2005. Cancer Epidemiol Biomark Prev. 2005, 14 (1): 243-250.
4.
go back to reference Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010, 127 (12): 2893-2917. 10.1002/ijc.25516.CrossRefPubMed Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010, 127 (12): 2893-2917. 10.1002/ijc.25516.CrossRefPubMed
5.
go back to reference Stefani G, Slack FJ: Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol. 2008, 9 (3): 219-230. 10.1038/nrm2347.CrossRefPubMed Stefani G, Slack FJ: Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol. 2008, 9 (3): 219-230. 10.1038/nrm2347.CrossRefPubMed
6.
go back to reference Fabian MR, Sonenberg N, Filipowicz W: Regulation of mRNA translation and stability by microRNAs. Annu Rev Biochem. 2010, 79: 351-379. 10.1146/annurev-biochem-060308-103103.CrossRefPubMed Fabian MR, Sonenberg N, Filipowicz W: Regulation of mRNA translation and stability by microRNAs. Annu Rev Biochem. 2010, 79: 351-379. 10.1146/annurev-biochem-060308-103103.CrossRefPubMed
7.
go back to reference Zhao J-J, Yang J, Lin J, Yao N, Zhu Y, Zheng J, Xu J, Cheng JQ, Lin J-Y, Ma X: Identification of miRNAs associated with tumorigenesis of retinoblastoma by miRNA microarray analysis. Childs Nerv Syst. 2009, 25 (1): 13-20. 10.1007/s00381-008-0701-x.CrossRefPubMed Zhao J-J, Yang J, Lin J, Yao N, Zhu Y, Zheng J, Xu J, Cheng JQ, Lin J-Y, Ma X: Identification of miRNAs associated with tumorigenesis of retinoblastoma by miRNA microarray analysis. Childs Nerv Syst. 2009, 25 (1): 13-20. 10.1007/s00381-008-0701-x.CrossRefPubMed
8.
go back to reference Hassan O, Ahmad A, Sethi S, Sarkar FH: Recent updates on the role of microRNAs in prostate cancer. J Hematol Oncol. 2012, 5 (9): 13- Hassan O, Ahmad A, Sethi S, Sarkar FH: Recent updates on the role of microRNAs in prostate cancer. J Hematol Oncol. 2012, 5 (9): 13-
9.
go back to reference Du L, Pertsemlidis A: microRNA regulation of cell viability and drug sensitivity in lung cancer. Expert Opin Biol Ther. 2012, 12 (9): 1221-1239. 10.1517/14712598.2012.697149.CrossRefPubMed Du L, Pertsemlidis A: microRNA regulation of cell viability and drug sensitivity in lung cancer. Expert Opin Biol Ther. 2012, 12 (9): 1221-1239. 10.1517/14712598.2012.697149.CrossRefPubMed
11.
go back to reference Acunzo M, Visone R, Romano G, Veronese A, Lovat F, Palmieri D, Bottoni A, Garofalo M, Gasparini P, Condorelli G: miR-130a targets MET and induces TRAIL-sensitivity in NSCLC by downregulating miR-221 and 222. Oncogene. 2011, 31 (5): 634-642.PubMedPubMedCentral Acunzo M, Visone R, Romano G, Veronese A, Lovat F, Palmieri D, Bottoni A, Garofalo M, Gasparini P, Condorelli G: miR-130a targets MET and induces TRAIL-sensitivity in NSCLC by downregulating miR-221 and 222. Oncogene. 2011, 31 (5): 634-642.PubMedPubMedCentral
12.
go back to reference Jeon HS, Lee SY, Lee EJ, Yun SC, Cha EJ, Choi E, Na MJ, Park JY, Kang J, Son JW: Combining microRNA-449a/b with a HDAC inhibitor has a synergistic effect on growth arrest in lung cancer. Lung Cancer. 2012, 76 (2): 171-176. 10.1016/j.lungcan.2011.10.012.CrossRefPubMed Jeon HS, Lee SY, Lee EJ, Yun SC, Cha EJ, Choi E, Na MJ, Park JY, Kang J, Son JW: Combining microRNA-449a/b with a HDAC inhibitor has a synergistic effect on growth arrest in lung cancer. Lung Cancer. 2012, 76 (2): 171-176. 10.1016/j.lungcan.2011.10.012.CrossRefPubMed
13.
go back to reference Zhang J-G, Wang J-J, Zhao F, Liu Q, Jiang K, Yang G-H: MicroRNA-21 (miR-21) represses tumor suppressor PTEN and promotes growth and invasion in non-small cell lung cancer (NSCLC). Clin Chim Acta. 2010, 411 (11): 846-852.CrossRefPubMed Zhang J-G, Wang J-J, Zhao F, Liu Q, Jiang K, Yang G-H: MicroRNA-21 (miR-21) represses tumor suppressor PTEN and promotes growth and invasion in non-small cell lung cancer (NSCLC). Clin Chim Acta. 2010, 411 (11): 846-852.CrossRefPubMed
14.
go back to reference Lebanony D, Benjamin H, Gilad S, Ezagouri M, Dov A, Ashkenazi K, Gefen N, Izraeli S, Rechavi G, Pass H: Diagnostic assay based on hsa-miR-205 expression distinguishes squamous from nonsquamous non-small-cell lung carcinoma. J Clin Oncol. 2009, 27 (12): 2030-2037. 10.1200/JCO.2008.19.4134.CrossRefPubMed Lebanony D, Benjamin H, Gilad S, Ezagouri M, Dov A, Ashkenazi K, Gefen N, Izraeli S, Rechavi G, Pass H: Diagnostic assay based on hsa-miR-205 expression distinguishes squamous from nonsquamous non-small-cell lung carcinoma. J Clin Oncol. 2009, 27 (12): 2030-2037. 10.1200/JCO.2008.19.4134.CrossRefPubMed
15.
go back to reference Liu X, Sempere LF, Guo Y, Korc M, Kauppinen S, Freemantle SJ, Dmitrovsky E: Involvement of microRNAs in lung cancer biology and therapy. Transl Res. 2011, 157 (4): 200-208. 10.1016/j.trsl.2011.01.001.CrossRefPubMedPubMedCentral Liu X, Sempere LF, Guo Y, Korc M, Kauppinen S, Freemantle SJ, Dmitrovsky E: Involvement of microRNAs in lung cancer biology and therapy. Transl Res. 2011, 157 (4): 200-208. 10.1016/j.trsl.2011.01.001.CrossRefPubMedPubMedCentral
16.
go back to reference Gao W, Yu Y, Cao H, Shen H, Li X, Pan S, Shu Y: Deregulated expression of miR-21, miR-143 and miR-181a in non small cell lung cancer is related to clinicopathologic characteristics or patient prognosis. Biomed Pharmacother. 2010, 64 (6): 399-408. 10.1016/j.biopha.2010.01.018.CrossRefPubMed Gao W, Yu Y, Cao H, Shen H, Li X, Pan S, Shu Y: Deregulated expression of miR-21, miR-143 and miR-181a in non small cell lung cancer is related to clinicopathologic characteristics or patient prognosis. Biomed Pharmacother. 2010, 64 (6): 399-408. 10.1016/j.biopha.2010.01.018.CrossRefPubMed
17.
go back to reference Yu F, Deng H, Yao H, Liu Q, Su F, Song E: Mir-30 reduction maintains self-renewal and inhibits apoptosis in breast tumor-initiating cells. Oncogene. 2010, 29 (29): 4194-4204. 10.1038/onc.2010.167.CrossRefPubMed Yu F, Deng H, Yao H, Liu Q, Su F, Song E: Mir-30 reduction maintains self-renewal and inhibits apoptosis in breast tumor-initiating cells. Oncogene. 2010, 29 (29): 4194-4204. 10.1038/onc.2010.167.CrossRefPubMed
18.
go back to reference Feber A, Wilson GA, Zhang L, Presneau N, Idowu B, Down TA, Rakyan VK, Noon LA, Lloyd AC, Stupka E: Comparative methylome analysis of benign and malignant peripheral nerve sheath tumors. Genome Res. 2011, 21 (4): 515-524. 10.1101/gr.109678.110.CrossRefPubMedPubMedCentral Feber A, Wilson GA, Zhang L, Presneau N, Idowu B, Down TA, Rakyan VK, Noon LA, Lloyd AC, Stupka E: Comparative methylome analysis of benign and malignant peripheral nerve sheath tumors. Genome Res. 2011, 21 (4): 515-524. 10.1101/gr.109678.110.CrossRefPubMedPubMedCentral
19.
go back to reference Quintavalle C, Donnarumma E, Iaboni M, Roscigno G, Garofalo M, Romano G, Fiore D, De Marinis P, Croce CM, Condorelli G: Effect of miR-21 and miR-30b/c on TRAIL-induced apoptosis in glioma cells. Oncogene. 2012, 31: 4001-4008.CrossRef Quintavalle C, Donnarumma E, Iaboni M, Roscigno G, Garofalo M, Romano G, Fiore D, De Marinis P, Croce CM, Condorelli G: Effect of miR-21 and miR-30b/c on TRAIL-induced apoptosis in glioma cells. Oncogene. 2012, 31: 4001-4008.CrossRef
20.
go back to reference Guan P, Yin Z, Li X, Wu W, Zhou B: Meta-analysis of human lung cancer microRNA expression profiling studies comparing cancer tissues with normal tissues. J Exp Clin Cancer Res. 2012, 31 (1): 54-10.1186/1756-9966-31-54.CrossRefPubMedPubMedCentral Guan P, Yin Z, Li X, Wu W, Zhou B: Meta-analysis of human lung cancer microRNA expression profiling studies comparing cancer tissues with normal tissues. J Exp Clin Cancer Res. 2012, 31 (1): 54-10.1186/1756-9966-31-54.CrossRefPubMedPubMedCentral
21.
go back to reference Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR: Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005, 33 (20): 179-10.1093/nar/gni178.CrossRef Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR: Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005, 33 (20): 179-10.1093/nar/gni178.CrossRef
22.
go back to reference Li J, Kong X, Zhang J, Luo Q, Li X, Fang L: MiRNA-26b inhibits proliferation by targeting PTGS2 in breast cancer. Cancer Cell Int. 2013, 13 (1): 7-10.1186/1475-2867-13-7.CrossRefPubMedPubMedCentral Li J, Kong X, Zhang J, Luo Q, Li X, Fang L: MiRNA-26b inhibits proliferation by targeting PTGS2 in breast cancer. Cancer Cell Int. 2013, 13 (1): 7-10.1186/1475-2867-13-7.CrossRefPubMedPubMedCentral
23.
go back to reference You X, Liu F, Zhang T, Li Y, Ye L, Zhang X: Hepatitis B virus X protein upregulates oncogene Rab18 to result in the dysregulation of lipogenesis and proliferation of hepatoma cells. Carcinogenesis. 2013, 34 (7): 1644-1652. 10.1093/carcin/bgt089.CrossRefPubMed You X, Liu F, Zhang T, Li Y, Ye L, Zhang X: Hepatitis B virus X protein upregulates oncogene Rab18 to result in the dysregulation of lipogenesis and proliferation of hepatoma cells. Carcinogenesis. 2013, 34 (7): 1644-1652. 10.1093/carcin/bgt089.CrossRefPubMed
24.
go back to reference Helwak A, Kudla G, Dudnakova T, Tollervey D: Mapping the human miRNA interactome by CLASH reveals frequent noncanonical binding. Cell. 2013, 153 (3): 654-665. 10.1016/j.cell.2013.03.043.CrossRefPubMedPubMedCentral Helwak A, Kudla G, Dudnakova T, Tollervey D: Mapping the human miRNA interactome by CLASH reveals frequent noncanonical binding. Cell. 2013, 153 (3): 654-665. 10.1016/j.cell.2013.03.043.CrossRefPubMedPubMedCentral
25.
go back to reference Liang Y: An expression meta-analysis of predicted microRNA targets identifies a diagnostic signature for lung cancer. BMC Med Genet. 2008, 1 (1): 61- Liang Y: An expression meta-analysis of predicted microRNA targets identifies a diagnostic signature for lung cancer. BMC Med Genet. 2008, 1 (1): 61-
26.
go back to reference Ouzounova M, Vuong T, Ancey P-B, Ferrand M, Durand G, Kelm FL-C, Croce C, Matar C, Herceg Z, Hernandez-Vargas H: MicroRNA miR-30 family regulates non-attachment growth of breast cancer cells. BMC Genomics. 2013, 14 (1): 139-10.1186/1471-2164-14-139.CrossRefPubMedPubMedCentral Ouzounova M, Vuong T, Ancey P-B, Ferrand M, Durand G, Kelm FL-C, Croce C, Matar C, Herceg Z, Hernandez-Vargas H: MicroRNA miR-30 family regulates non-attachment growth of breast cancer cells. BMC Genomics. 2013, 14 (1): 139-10.1186/1471-2164-14-139.CrossRefPubMedPubMedCentral
28.
go back to reference Martinez I, Cazalla D, Almstead LL, Steitz JA, DiMaio D: miR-29 and miR-30 regulate B-Myb expression during cellular senescence. Proc Natl Acad Sci. 2011, 108 (2): 522-527. 10.1073/pnas.1017346108.CrossRefPubMed Martinez I, Cazalla D, Almstead LL, Steitz JA, DiMaio D: miR-29 and miR-30 regulate B-Myb expression during cellular senescence. Proc Natl Acad Sci. 2011, 108 (2): 522-527. 10.1073/pnas.1017346108.CrossRefPubMed
29.
go back to reference Chia WJ, Tang BL: Emerging roles for Rab family GTPases in human cancer. Biochim Biophys Acta. 2009, 1795 (2): 110-116.PubMed Chia WJ, Tang BL: Emerging roles for Rab family GTPases in human cancer. Biochim Biophys Acta. 2009, 1795 (2): 110-116.PubMed
30.
go back to reference Tang BL, Ng EL: Rabs and cancer cell motility. Cell Motil Cytoskeleton. 2009, 66 (7): 365-370. 10.1002/cm.20376.CrossRefPubMed Tang BL, Ng EL: Rabs and cancer cell motility. Cell Motil Cytoskeleton. 2009, 66 (7): 365-370. 10.1002/cm.20376.CrossRefPubMed
31.
go back to reference Cheng KW, Lahad JP, Gray JW, Mills GB: Emerging role of RAB GTPases in cancer and human disease. Cancer Res. 2005, 65 (7): 2516-2519. 10.1158/0008-5472.CAN-05-0573.CrossRefPubMed Cheng KW, Lahad JP, Gray JW, Mills GB: Emerging role of RAB GTPases in cancer and human disease. Cancer Res. 2005, 65 (7): 2516-2519. 10.1158/0008-5472.CAN-05-0573.CrossRefPubMed
32.
go back to reference Bem D, Yoshimura S-I, Nunes-Bastos R, Bond FF, Kurian MA, Rahman F, Handley MTW, Hadzhiev Y, Masood I, Straatman-Iwanowska AA: Loss-of-function mutations in RAB18 cause Warburg micro syndrome. Am J Hum Genet. 2011, 88 (4): 499-507. 10.1016/j.ajhg.2011.03.012.CrossRefPubMedPubMedCentral Bem D, Yoshimura S-I, Nunes-Bastos R, Bond FF, Kurian MA, Rahman F, Handley MTW, Hadzhiev Y, Masood I, Straatman-Iwanowska AA: Loss-of-function mutations in RAB18 cause Warburg micro syndrome. Am J Hum Genet. 2011, 88 (4): 499-507. 10.1016/j.ajhg.2011.03.012.CrossRefPubMedPubMedCentral
33.
go back to reference Behrends U, Schneider I, Rössler S, Frauenknecht H, Golbeck A, Lechner B, Eigenstetter G, Zobywalski C, Müller-Weihrich S, Graubner U: Novel tumor antigens identified by autologous antibody screening of childhood medulloblastoma cDNA libraries. Int J Cancer. 2003, 106 (2): 244-251. 10.1002/ijc.11208.CrossRefPubMed Behrends U, Schneider I, Rössler S, Frauenknecht H, Golbeck A, Lechner B, Eigenstetter G, Zobywalski C, Müller-Weihrich S, Graubner U: Novel tumor antigens identified by autologous antibody screening of childhood medulloblastoma cDNA libraries. Int J Cancer. 2003, 106 (2): 244-251. 10.1002/ijc.11208.CrossRefPubMed
Metadata
Title
microRNA-30b/c inhibits non-small cell lung cancer cell proliferation by targeting Rab18
Authors
Keng Zhong
Kun Chen
Lin Han
Bailing Li
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2014
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-14-703

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