Inhibition mechanism of lung cancer cell metastasis through targeted regulation of Smad3 by miR‑15a

Guo,Shuai; Li,Ming; Li,Juan; Lv,Yan

doi:10.3892/ol.2019.11194

Inhibition mechanism of lung cancer cell metastasis through targeted regulation of Smad3 by miR‑15a

Authors:
- Shuai Guo
- Ming Li
- Juan Li
- Yan Lv
View Affiliations

Affiliations: Department of Medical Oncology, Shandong Provincial Chest Hospital, Jinan, Shandong 250013, P.R. China, Department of Thoracic Surgery, Shandong Provincial Chest Hospital, Jinan, Shandong 250013, P.R. China, Department of Pathology, The Forth Hospital of Jinan, Jinan, Shandong 250031, P.R. China, Department of Internal Medicine Ward IV, Shandong Provincial Chest Hospital, Jinan, Shandong 250013, P.R. China
Published online on: December 10, 2019 https://doi.org/10.3892/ol.2019.11194
Pages: 1516-1522
Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Effect of targeted regulation of mothers against decapentaplegic homolog 3 (Smad3) by microRNA-15a (miR‑15a) on the proliferation, invasion and metastasis of non‑small cell lung cancer (NSCLC) cells and its related mechanisms were investigated. Fifty pairs of NSCLC and para‑cancerous tissues were collected to identify the expression level of miR‑15a in NSCLC, para‑cancerous tissue, and cell lines A549, H1299, H1975 and BEAS‑2B by real‑time fluorescence quantitative PCR (RT‑PCR); A549 cells were transfected with miR‑15a mimic; the MTT assay was performed to detect the role of miR‑15a transfection in proliferation of A549 cells, the wound healing assay was carried out to identify the role of miR‑15a in migration of A549 cells; Transwell invasion assay was conducted to analyze the role of miR‑15a in invasion of A549 cells; western blotting was carried out to find the effect of miR‑15a on Smad3 expression, and Spearman's rank correlation was used to analyze the correlation between miR‑15a and Smad3 expression. NSCLC tissues and cells showed significantly lower miR‑15a expression, compared with para‑cancerous tissues and normal cell lines (P=0.023). miR‑15a was significantly more expressed in A549 cells transfected with miR‑15a mimic (P=0.043). Overexpression of miR‑15a can significantly inhibit A549 cell proliferation (P=0.038), migration (P=0.033) and invasion (P=0.025), and significantly reduced the expression level of Smad3 (P=0.031) in A549 cells. Spearman's rank correlation showed negative correlation of miR‑15a expression with Smad3, which may indicate negative regulation (r=‑0.34, P<0.0001). Inhibition of proliferation, migration and invasion of NSCLC cells can be achieved with targeted regulation of Smad3 by miR‑15a.

View Figures

View References

1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
2	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2019. CA Cancer J Clin. 69:7–34. 2019. View Article : Google Scholar : PubMed/NCBI
3	van Klaveren RJ: Lung cancer screening. Eur J Cancer. 47 (Suppl 3):S147–S155. 2011. View Article : Google Scholar : PubMed/NCBI
4	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
5	Bouyssou JM, Manier S, Huynh D, Issa S, Roccaro AM and Ghobrial IM: Regulation of microRNAs in cancer metastasis. Biochim Biophys Acta. 1845:255–265. 2014.PubMed/NCBI
6	Zagryazhskaya A and Zhivotovsky B: miRNAs in lung cancer: A link to aging. Ageing Res Rev. 17:54–67. 2014. View Article : Google Scholar : PubMed/NCBI
7	Wan L, Zhang L, Fan K and Wang J: miR-27b targets LIMK1 to inhibit growth and invasion of NSCLC cells. Mol Cell Biochem. 390:85–91. 2014. View Article : Google Scholar : PubMed/NCBI
8	Zhan M, Qu Q, Wang G and Zhou H: Let-7c sensitizes acquired cisplatin-resistant A549 cells by targeting ABCC2 and Bcl-XL. Pharmazie. 68:955–961. 2013.PubMed/NCBI
9	Zhang N, Wei X and Xu L: miR-150 promotes the proliferation of lung cancer cells by targeting P53. FEBS Lett. 587:2346–2351. 2013. View Article : Google Scholar : PubMed/NCBI
10	Wu Y, Huang J, Xu H and Gong Z: Over-expression of miR-15a-3p enhances the radiosensitivity of cervical cancer by targeting tumor protein D52. Biomed Pharmacother. 105:1325–1334. 2018. View Article : Google Scholar : PubMed/NCBI
11	Jin J, Zhang J, Xue Y, Luo L, Wang S and Tian H: miRNA-15a regulates the proliferation and apoptosis of papillary thyroid carcinoma via regulating AKT pathway. OncoTargets Ther. 12:6217–6226. 2019. View Article : Google Scholar
12	Molina-Pinelo S, Gutiérrez G, Pastor MD, Hergueta M, Moreno-Bueno G, García-Carbonero R, Nogal A, Suárez R, Salinas A, Pozo-Rodríguez F, et al: MicroRNA-dependent regulation of transcription in non-small cell lung cancer. PLoS One. 9:e905242014. View Article : Google Scholar : PubMed/NCBI
13	O'Connell RM, Rao DS, Chaudhuri AA and Baltimore D: Physiological and pathological roles for microRNAs in the immune system. Nat Rev Immunol. 10:111–122. 2010. View Article : Google Scholar : PubMed/NCBI
14	Codony C, Crespo M, Abrisqueta P, Montserrat E and Bosch F: Gene expression profiling in chronic lymphocytic leukaemia. Best Pract Res Clin Haematol. 22:211–222. 2009. View Article : Google Scholar : PubMed/NCBI
15	Pekarsky Y and Croce CM: Role of miR-15/16 in CLL. Cell Death Differ. 22:6–11. 2015. View Article : Google Scholar : PubMed/NCBI
16	Aqeilan RI, Calin GA and Croce CM: miR-15a and miR-16-1 in cancer: Discovery, function and future perspectives. Cell Death Differ. 17:215–220. 2010. View Article : Google Scholar : PubMed/NCBI
17	Musumeci M, Coppola V, Addario A, Patrizii M, Maugeri-Saccà M, Memeo L, Colarossi C, Francescangeli F, Biffoni M, Collura D, et al: Control of tumor and microenvironment cross-talk by miR-15a and miR-16 in prostate cancer. Oncogene. 30:4231–4242. 2011. View Article : Google Scholar : PubMed/NCBI
18	Jin W, Chen F, Wang K, Song Y, Fei X and Wu B: miR-15a/miR-16 cluster inhibits invasion of prostate cancer cells by suppressing TGF-β signaling pathway. Biomed Pharmacother. 104:637–644. 2018. View Article : Google Scholar : PubMed/NCBI
19	Bonci D, Coppola V, Musumeci M, Addario A, Giuffrida R, Memeo L, D'Urso L, Pagliuca A, Biffoni M, Labbaye C, et al: The miR-15a-miR-16-1 cluster controls prostate cancer by targeting multiple oncogenic activities. Nat Med. 14:1271–1277. 2008. View Article : Google Scholar : PubMed/NCBI
20	Janaki Ramaiah M, Lavanya A, Honarpisheh M, Zarea M, Bhadra U and Bhadra MP: miR-15/16 complex targets p70S6 kinase 1 and controls cell proliferation in MDA-MB-231 breast cancer cells. Gene. 552:255–264. 2014. View Article : Google Scholar : PubMed/NCBI
21	Pouliot LM, Chen YC, Bai J, Guha R, Martin SE, Gottesman MM and Hall MD: Cisplatin sensitivity mediated by WEE1 and CHK1 is mediated by miR-155 and the miR-15 family. Cancer Res. 72:5945–5955. 2012. View Article : Google Scholar : PubMed/NCBI
22	Bozok Çetintaş V, Tetik Vardarlı A, Düzgün Z, Tezcanlı Kaymaz B, Açıkgöz E, Aktuğ H, Kosova Can B, Gündüz C and Eroğlu Z: miR-15a enhances the anticancer effects of cisplatin in the resistant non-small cell lung cancer cells. Tumour Biol. 37:1739–1751. 2016. View Article : Google Scholar : PubMed/NCBI
23	Çalışkan M, Güler H and Bozok Çetintaş V: Current updates on microRNAs as regulators of chemoresistance. Biomed Pharmacother. 95:1000–1012. 2017. View Article : Google Scholar : PubMed/NCBI
24	Renjie W and Haiqian L: miR-132, miR-15a and miR-16 synergistically inhibit pituitary tumor cell proliferation, invasion and migration by targeting Sox5. Cancer Lett. 356B:568–578. 2015. View Article : Google Scholar
25	Shi L, Jackstadt R, Siemens H, Li H, Kirchner T and Hermeking H: p53-induced miR-15a/16-1 and AP4 form a double-negative feedback loop to regulate epithelial-mesenchymal transition and metastasis in colorectal cancer. Cancer Res. 74:532–542. 2014. View Article : Google Scholar : PubMed/NCBI
26	Gao W, Wang Y, Wang W and Shi L: The first multiplication atom-bond connectivity index of molecular structures in drugs. Saudi Pharm J. 25:548–555. 2017. View Article : Google Scholar : PubMed/NCBI
27	He J: Knocking down miR-15a expression promotes the occurrence and development and induces the EMT of NSCLC cells in vitro. Saudi J Biol Sci. 24:1859–1865. 2017. View Article : Google Scholar : PubMed/NCBI
28	Li H, Xu D, Toh BH and Liu JP: TGF-beta and cancer: Is Smad3 a repressor of hTERT gene? Cell Res. 16:169–173. 2006. View Article : Google Scholar : PubMed/NCBI
29	Xu W, Zeng F, Li S, Li G, Lai X, Wang QJ and Deng F: Crosstalk of protein kinase C ε with Smad2/3 promotes tumor cell proliferation in prostate cancer cells by enhancing aerobic glycolysis. Cell Mol Life Sci. 75:4583–4598. 2018. View Article : Google Scholar : PubMed/NCBI
30	Paul D, Dixit A, Srivastava A, Tripathi M, Prakash D, Sarkar C, Ramanujam B, Banerjee J and Chandra PS: Altered transforming growth factor beta/SMAD3 signalling in patients with hippocampal sclerosis. Epilepsy Res. 146:144–150. 2018. View Article : Google Scholar : PubMed/NCBI
31	Ooshima A, Park J and Kim SJ: Phosphorylation status at Smad3 linker region modulates transforming growth factor-β-induced epithelial-mesenchymal transition and cancer progression. Cancer Sci. 110:481–488. 2019. View Article : Google Scholar : PubMed/NCBI
32	Wang Y, Xiang J, Wang J and Ji Y: Downregulation of TGF-β1 suppressed proliferation and increased chemosensitivity of ovarian cancer cells by promoting BRCA1/Smad3 signaling. Biol Res. 51:582018. View Article : Google Scholar : PubMed/NCBI