Effect of miR‑140‑5p on the regulation of proliferation and apoptosis in NSCLC and its underlying mechanism

Zhou,Wenwen; Wang,Xiaoyu; Yin,Duanduan; Xue,Lei; Ma,Zhongfeng; Wang,Zhenzhen; Zhang,Qianyi; Zhao,Zishu; Wang,Haixia; Sun,Yan; Yang,Yanhong

doi:10.3892/etm.2019.7701

Effect of miR‑140‑5p on the regulation of proliferation and apoptosis in NSCLC and its underlying mechanism

Authors:
- Wenwen Zhou
- Xiaoyu Wang
- Duanduan Yin
- Lei Xue
- Zhongfeng Ma
- Zhenzhen Wang
- Qianyi Zhang
- Zishu Zhao
- Haixia Wang
- Yan Sun
- Yanhong Yang
View Affiliations

Affiliations: Department of Oncology, Qinhuangdao First People's Hospital, Qinhuangdao, Hebei 066000, P.R. China, Foundation and Clinic of Malignant Tumor, Postgraduate College, Chengde Medical University, Chengde, Hebei 067000, P.R. China, Division of Pharmacy, University of Tasmania, Hobart, Tasmania 7005, Australia, Foundation and Clinic of Malignant Tumor, Postgraduate College, Hebei Medical University, Shijiazhuang, Hebei 051117, P.R. China
Published online on: June 21, 2019 https://doi.org/10.3892/etm.2019.7701
Pages: 1350-1356

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

Non‑small cell lung cancer (NSCLC) is the most common type of lung cancer accounting for ~80% of lung cancer cases. According to novel research, numerous microRNAs (miRs) have been suggested to function as important regulators of cancer. In addition, the expression of miR‑140‑5p is decreased in patients with NSCLC. Therefore, it is important to further elucidate the role of miR‑140‑5p in NSCLC. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was used in order to investigate the expression of miR‑140‑5p in NSCLC tissues and matched normal tissues and to determine miR‑140‑5p levels following transfection with mimics into A549 lung cancer cells. Targetscan software was used to predict the oncogene target of miR‑140‑5p. This analysis revealed that YES proto‑oncogene 1 (YES1) includes a target site for miR‑140‑5p binding. The results revealed that YES1 is a potential target gene of miR‑140‑5p, and this was further confirmed by the results of luciferase reporter assays, which demonstrated that miR‑140‑5p directly targeted the predicted binding site in the 3'‑untranslated region of YES1. Cell Counting Kit‑8 (CCK‑8) and flow cytometry assays were performed to determine the levels of cell viability and apoptosis. Western blot assays was performed to investigate the expression levels of YES1 and proteins associated with apoptosis in A549 cells following transfection. The results revealed that miR‑140‑5p expression was significantly downregulated in NSCLC tissues compared with matched normal tissues. The expression of miR‑140‑5p was significantly increased following transfection with miR‑140‑5p mimics. The results of CCK‑8 and flow cytometry assays indicated that miR‑140‑5p inhibited proliferation and induced apoptosis of tumor cells. Western blot analysis and RT‑qPCR revealed that YES1 and B‑cell lymphoma 2 (Bcl‑2) mRNA and protein expression levels were markedly decreased in A549 cells, while Bcl‑2 associated X (Bax) and caspase‑3 expression levels increased significantly following transfection with miR‑140‑5p mimics compared with the negative control group. In conclusion, miR‑140‑5p may induce apoptosis in A549 cells by targeting YES1 and regulating the expression of apoptosis‑associated proteins Bcl‑2, Bax and caspase‑3.

View Figures

View References

1	Liu P, Pu J, Zhang J, Chen Z, Wei K and Shi L: Bioinformatic analysis of miR-4792 regulates Radix Tetrastigma hemsleyani flavone to inhibit proliferation, invasion, and induce apoptosis of A549 cells. Onco Targets Ther. 12:1401–1412. 2019. View Article : Google Scholar : PubMed/NCBI
2	Pope CA III, Burnett RT, Thun MJ, Calle EE, Krewski D, Ito K and Thurston GD: Lung cancer, cardiopulmonary mortality, and long-term exposure to fine particulate air pollution. JAMA. 287:1132–1141. 2002. View Article : Google Scholar : PubMed/NCBI
3	Sui A, Zhang X and Zhu Q: Diagnostic value of serum miR197 and miR145 in non-small cell lung cancer. Oncol Lett. 17:3247–3252. 2019.PubMed/NCBI
4	Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, et al: Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 350:2129–2139. 2004. View Article : Google Scholar : PubMed/NCBI
5	Carvalho PE, Antonãngelo L, Bernardi FD, Leão LE, Rodrigues OR and Capelozzi VL: Useful prognostic panel markers to express the biological tumor status in resected lung adenocarcinomas. Jpn J Clin Oncol. 30:478–486. 2000. View Article : Google Scholar : PubMed/NCBI
6	Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI
8	Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ and Lötvall JO: Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 9:654–659. 2007. View Article : Google Scholar : PubMed/NCBI
9	He L and Hannon GJ: MicroRNAs: Small RNAs with a big role in gene regulation. Nat Rev Genet. 5:522–531. 2004. View Article : Google Scholar : PubMed/NCBI
10	Taylor DD and Gercel-Taylor C: MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol. 110:13–21. 2008. View Article : Google Scholar : PubMed/NCBI
11	Dews M, Homayouni A, Yu D, Murphy D, Sevignani C, Wentzel E, Furth EE, Lee WM, Enders GH, Mendell JT and Thomas-Tikhonenko A: Augmentation of tumor angiogenesis by a Myc-activated microRNA cluster. Nat Genet. 38:1060–1065. 2006. View Article : Google Scholar : PubMed/NCBI
12	Ladeiro Y, Couchy G, Balabaud C, Bioulac-Sage P, Pelletier L, Rebouissou S and Zucman-Rossi J: MicroRNA profiling in hepatocellular tumors is associated with clinical features and oncogene/tumor suppressor gene mutations. Hepatology. 47:1955–1963. 2008. View Article : Google Scholar : PubMed/NCBI
13	Mitra R, Edmonds MD, Sun J, Zhao M, Yu H, Eischen CM and Zhao Z: Reproducible combinatorial regulatory networks elucidate novel oncogenic microRNAs in non-small cell lung cancer. RNA. 20:1356–1368. 2014. View Article : Google Scholar : PubMed/NCBI
14	Wozniak MB, Scelo G, Muller DC, Mukeria A, Zaridze D and Brennan P: Circulating microRNAs as non-invasive biomarkers for early detection of non-small-cell lung cancer. PLoS One. 10:e01250262015. View Article : Google Scholar : PubMed/NCBI
15	Yao Q, Zhang AM, Ma H, Lin S, Wang XX, Sun JG and Chen ZT: Novel molecular beacons to monitor microRNAs in non-small-cell lung cancer. Mol Cell Probes. 26:182–187. 2012. View Article : Google Scholar : PubMed/NCBI
16	Goga A and Benz C: Anti-oncomir suppression of tumor phenotypes. Mol Interv. 7:199–202, 180. 2007. View Article : Google Scholar : PubMed/NCBI
17	Yan X, Zhu Z, Xu S, Yang LN, Liao XH, Zheng M, Yang D, Wang J, Chen D, Wang L, et al: MicroRNA-140-5p inhibits hepatocellular carcinoma by directly targeting the unique isomerase Pin1 to block multiple cancer-driving pathways. Sci Rep. 7:459152017. View Article : Google Scholar : PubMed/NCBI
18	Zhou CW, Zhao WJ, Zhu YG and Zhao XD: MiR-185 inhibits tumor growth and enhances chemo-resistance via targeting SRY-related high mobility group box transcription factor 13 in non-small-cell carcinoma. Am J Transl Res. 10:2600–2609. 2018.PubMed/NCBI
19	Yang H, Fang F, Chang R and Yang L: MicroRNA-140-5p suppresses tumor growth and metastasis by targeting transforming growth factor β receptor 1 and fibroblast growth factor 9 in hepatocellular carcinoma. Hepatology. 58:205–217. 2013. View Article : Google Scholar : PubMed/NCBI
20	Zhang W, Zou C, Pan L, Xu Y, Qi W, Ma G, Hou Y and Jiang P: MicroRNA-140-5p inhibits the progression of colorectal cancer by targeting VEGFA. Cell Physiol Biochem. 37:1123–1133. 2015. View Article : Google Scholar : PubMed/NCBI
21	Dong C, Liu X, Wang H, Li J, Dai L, Li J and Xu Z: Hypoxic non-small-cell lung cancer cell-derived exosomal miR-21 promotes resistance of normoxic cell to cisplatin. Onco Targets Ther. 12:1947–1956. 2019. View Article : Google Scholar : PubMed/NCBI
22	Flamini V, Jiang WG and Cui Y: Therapeutic Role of MiR-140-5p for the treatment of non-small cell lung cancer. Anticancer Res. 37:4319–4327. 2017.PubMed/NCBI
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Arcà B, Colantoni A, Fiorillo C, Severini F, Benes V, Di Luca M, Calogero RA and Lombardo F: MicroRNAs from saliva of anopheline mosquitoes mimic human endogenous miRNAs and may contribute to vector-host-pathogen interactions. Sci Rep. 9:29552019. View Article : Google Scholar : PubMed/NCBI
25	Mirzaei H, Khataminfar S, Mohammadparast S, Sales SS, Maftouh M, Mohammadi M, Simonian M, Parizadeh SM, Hassanian SM and Avan A: Circulating microRNAs as potential diagnostic biomarkers and therapeutic targets in gastric cancer: Current status and future perspectives. Curr Med Chem. 23:4135–4150. 2016. View Article : Google Scholar : PubMed/NCBI
26	Long XB, Sun GB, Hu S, Liang GT, Wang N, Zhang XH, Cao PP, Zhen HT, Cui YH and Liu Z: Let-7a microRNA functions as a potential tumor suppressor in human laryngeal cancer. Oncol Rep. 22:1189–1195. 2009.PubMed/NCBI
27	Cheng TF, Choudhuri S and Muldoon-Jacobs K: Epigenetic targets of some toxicologically relevant metals: A review of the literature. J Appl Toxicol. 32:643–653. 2012. View Article : Google Scholar : PubMed/NCBI
28	Wang K, Chen M and Wu W: Analysis of microRNA (miRNA) expression profiles reveals 11 key biomarkers associated with non-small cell lung cancer. World J Surg Oncol. 15:1752017. View Article : Google Scholar : PubMed/NCBI
29	Hu L, Ai J, Long H, Liu W, Wang X, Zuo Y, Li Y, Wu Q and Deng Y: Integrative microRNA and gene profiling data analysis reveals novel biomarkers and mechanisms for lung cancer. Oncotarget. 7:8441–8454. 2016.PubMed/NCBI
30	Xiao X, Mruk DD, Lee WM and Cheng CY: c-Yes regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization in the seminiferous epithelium of rat testes. Int J Biochem Cell Biol. 43:651–665. 2011. View Article : Google Scholar : PubMed/NCBI
31	Bilal E, Alexe G, Yao M, Cong L, Kulkarni A, Ginjala V, Toppmeyer D, Ganesan S and Bhanot G: Identification of the YES1 kinase as a therapeutic target in basal-like breast cancers. Genes Cancer. 1:1063–1073. 2010. View Article : Google Scholar : PubMed/NCBI
32	Seki T, Fujii G, Mori S, Tamaoki N and Shibuya M: Amplification of c-yes-1 proto-oncogene in a primary human gastric cancer. Jpn J Cancer Res. 76:907–910. 1985.PubMed/NCBI
33	Liu L, Yang J, Zhu X, Li D, Lv Z and Zhang X: Long noncoding RNA H19 competitively binds miR-17-5p to regulate YES1 expression in thyroid cancer. FEBS J. 283:2326–2339. 2016. View Article : Google Scholar : PubMed/NCBI
34	Lee SA, Kim JS, Park SY, Kim HJ, Yu SK, Kim CS, Chun HS, Kim J, Park JT, Go D and Kim DK: miR-203 downregulates Yes-1 and suppresses oncogenic activity in human oral cancer cells. J Biosci Bioeng. 120:351–358. 2015. View Article : Google Scholar : PubMed/NCBI
35	Fang Z, Yin S, Sun R, Zhang S, Fu M, Wu Y, Zhang T, Khaliq J and Li Y: miR-140-5p suppresses the proliferation, migration and invasion of gastric cancer by regulating YES1. Mol Cancer. 16:1392017. View Article : Google Scholar : PubMed/NCBI
36	Mohamed MS, Bishr MK, Almutairi FM and Ali AG: Inhibitors of apoptosis: Clinical implications in cancer. Apoptosis. 22:1487–1509. 2017. View Article : Google Scholar : PubMed/NCBI
37	Warren CFA, Wong-Brown MW and Bowden NA: BCL-2 family isoforms in apoptosis and cancer. Cell Death Dis. 10:1772019. View Article : Google Scholar : PubMed/NCBI
38	Julien O and Wells JA: Caspases and their substrates. Cell Death Differ. 24:1380–1389. 2017. View Article : Google Scholar : PubMed/NCBI
39	Adams CM, Clark-Garvey S, Porcu P and Eischen CM: Targeting the Bcl-2 family in B cell lymphoma. Front Oncol. 8:6362019. View Article : Google Scholar : PubMed/NCBI
40	Lin L, Cheng K, Xie Z, Chen C, Chen L, Huang Y and Liang Z: Purification and characterization a polysaccharide from Hedyotis diffusa and its apoptosis inducing activity toward human lung cancer cell line A549. Int J Biol Macromol. 122:64–71. 2019. View Article : Google Scholar : PubMed/NCBI
41	Xu P, Cai X, Zhang W, Li Y, Qiu P, Lu D and He X: Flavonoids of Rosa roxburghii Tratt exhibit radioprotection and anti-apoptosis properties via the Bcl-2(Ca(2+))/Caspase-3/PARP-1 pathway. Apoptosis. 21:1125–1143. 2016. View Article : Google Scholar : PubMed/NCBI