MicroRNA-29a functions as a potential tumor suppressor through directly targeting CDC42 in non-small cell lung cancer

Li,Yongqiang; Wang,Zhi; Li,Yijiang; Jing,Ruijun

doi:10.3892/ol.2017.5888

MicroRNA-29a functions as a potential tumor suppressor through directly targeting CDC42 in non-small cell lung cancer

Authors:
- Yongqiang Li
- Zhi Wang
- Yijiang Li
- Ruijun Jing
View Affiliations

Affiliations: Department of Emergency, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710038, P.R. China, Department of Thoracic Surgery, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710038, P.R. China
Published online on: March 22, 2017 https://doi.org/10.3892/ol.2017.5888
Pages: 3896-3904

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

The expression and function of microRNA-29a (miR-29a) have been investigated in various types of cancer. In the present study, the expression, function and underlying molecular mechanism of miR‑29a were investigated in non‑small cell lung cancer (NSCLC). The expression level of miR-29a in NSCLC was determined using reverse transcription‑quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, migration and invasion ability were determined using Cell Counting Kit‑8, cell migration and invasion assays, respectively. Bioinformatics analysis and dual‑luciferase reporter assays were performed to determine whether cell division cycle 42 (CDC42) is a direct target gene of miR‑29a. To assess CDC42 mRNA and protein expression following transfection with miR‑29a, RT‑qPCR and western blotting were performed. Following knockdown of CDC42, functional assays were performed to investigate the roles of CDC42 in NSCLC. The results demonstrated that miR‑29a was downregulated in NSCLC and the decreased expression level of miR‑29a was significantly associated with advanced tumor‑node‑metastasis classification and metastasis. In addition, upregulation of miR‑29a inhibited cell proliferation, migration and invasion in NSCLC, whereas downregulation of miR‑29a had the opposite effects. Furthermore, CDC42 was identified as a direct target gene of miR‑29a in vitro. miR‑29a was demonstrated to function as a tumor suppressor in NSCLC by directly targeting CDC42 and may be investigated further as a target therapy for NSCLC.

View Figures

View References

1	Ilic N, Petricevic A, Arar D, Kotarac S, Banovic J, Ilic NF, Tripkovic A and Grandic L: Skip mediastinal nodal metastases in the IIIa/N2 non-small cell lung cancer. J Thorac Oncol. 2:1018–1021. 2007. View Article : Google Scholar : PubMed/NCBI
2	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
3	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2015. CA Cancer J Clin. 65:5–29. 2015. View Article : Google Scholar : PubMed/NCBI
4	Boffetta P and Nyberg F: Contribution of environmental factors to cancer risk. Br Med Bull. 68:71–94. 2003. View Article : Google Scholar : PubMed/NCBI
5	Didkowska J, Manczuk M, McNeill A, Powles J and Zatonski W: Lung cancer mortality at ages 35–54 in the European Union: Ecological study of evolving tobacco epidemics. BMJ. 331:189–191. 2005. View Article : Google Scholar : PubMed/NCBI
6	Ridge CA, McErlean AM and Ginsberg MS: Epidemiology of lung cancer. Semin Intervent Radiol. 30:93–98. 2013. View Article : Google Scholar : PubMed/NCBI
7	Paliogiannis P, Attene F, Cossu A, Budroni M, Cesaraccio R, Tanda F, Trignano M and Palmieri G: Lung cancer epidemiology in North Sardinia, Italy. Multidiscip Respir Med. 8:452013. View Article : Google Scholar : PubMed/NCBI
8	Ni T, Mao G, Xue Q, Liu Y, Chen B, Cui X, Lv L, Jia L, Wang Y and Ji L: Upregulated expression of ILF2 in non-small cell lung cancer is associated with tumor cell proliferation and poor prognosis. J Mol Histol. 46:325–335. 2015. View Article : Google Scholar : PubMed/NCBI
9	Peters S, Adjei AA, Gridelli C, Reck M, Kerr K and Felip E: ESMO Guidelines Working Group: Metastatic non-small-cell lung cancer (NSCLC): ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 23:(Suppl 7). vii56–vii64. 2012. View Article : Google Scholar : PubMed/NCBI
10	Zhang T, Zhang DM, Zhao D, Hou XM, Liu XJ, Ling XL and Ma SC: The prognostic value of osteopontin expression in non-small cell lung cancer: A meta-analysis. J Mol Histol. 45:533–540. 2014. View Article : Google Scholar : PubMed/NCBI
11	Cai J, Fang L, Huang Y, Li R, Yuan J, Yang Y, Zhu X, Chen B, Wu J and Li M: miR-205 targets PTEN and PHLPP2 to augment AKT signaling and drive malignant phenotypes in non-small cell lung cancer. Cancer Res. 73:5402–5415. 2013. View Article : Google Scholar : PubMed/NCBI
12	Pisters KM, Evans WK, Azzoli CG, Kris MG, Smith CA, Desch CE, Somerfield MR, Brouwers MC, Darling G, Ellis PM, et al: Cancer care ontario and American society of clinical oncology adjuvant chemotherapy and adjuvant radiation therapy for stages I-IIIA resectable non small-cell lung cancer guideline. J Clin Oncol. 25:5506–5518. 2007. View Article : Google Scholar : PubMed/NCBI
13	Verdecchia A, Francisci S, Brenner H, Gatta G, Micheli A, Mangone L and Kunkler I: EUROCARE-4 Working Group: Recent cancer survival in Europe: A 2000–02 period analysis of EUROCARE-4 data. Lancet Oncol. 8:784–796. 2007. View Article : Google Scholar : PubMed/NCBI
14	Sun W, Yuan X, Tian Y, Wu H, Xu H, Hu G and Wu K: Non-invasive approaches to monitor EGFR-TKI treatment in non-small-cell lung cancer. J Hematol Oncol. 8:952015. View Article : Google Scholar : PubMed/NCBI
15	Liu R, Liu X, Zheng Y, Gu J, Xiong S, Jiang P, Jiang X, Huang E, Yang Y, Ge D and Chu Y: MicroRNA-7 sensitizes non-small cell lung cancer cells to paclitaxel. Oncol Lett. 8:2193–2200. 2014.PubMed/NCBI
16	Ma Y, Li X, Cheng S, Wei W and Li Y: MicroRNA-106a confers cisplatin resistance in non-small cell lung cancer A549 cells by targeting adenosine triphosphatase-binding cassette A1. Mol Med Rep. 11:625–632. 2015.PubMed/NCBI
17	Zhong Z, Xia Y, Wang P, Liu B and Chen Y: Low expression of microRNA-30c promotes invasion by inducing epithelial mesenchymal transition in non-small cell lung cancer. Mol Med Rep. 10:2575–2579. 2014.PubMed/NCBI
18	Shi Y, Liu C, Liu X, Tang DG and Wang J: The microRNA miR-34a inhibits non-small cell lung cancer (NSCLC) growth and the CD44hi stem-like NSCLC cells. PLoS One. 9:e900222014. View Article : Google Scholar : PubMed/NCBI
19	Li X, Abdel-Mageed AB, Mondal D and Kandil E: MicroRNA expression profiles in differentiated thyroid cancer, a review. Int J Clin Exp Med. 6:74–80. 2013.PubMed/NCBI
20	Hwang HW and Mendell JT: MicroRNAs in cell proliferation, cell death, and tumorigenesis. Br J Cancer. 96:(Suppl). R40–R44. 2007.PubMed/NCBI
21	Tafsiri E, Darbouy M, Shadmehr MB, Zagryazhskaya A, Alizadeh J and Karimipoor M: Expression of miRNAs in non-small-cell lung carcinomas and their association with clinicopathological features. Tumour Biol. 36:1603–1612. 2015. View Article : Google Scholar : PubMed/NCBI
22	Braun J and Hüttelmaier S: Pathogenic mechanisms of deregulated microRNA expression in thyroid carcinomas of follicular origin. Thyroid Res. 4:(Suppl 1). S12011. View Article : Google Scholar : PubMed/NCBI
23	Zhang B, Pan X, Cobb GP and Anderson TA: microRNAs as oncogenes and tumor suppressors. Dev Biol. 302:1–12. 2007. View Article : Google Scholar : PubMed/NCBI
24	Esquela-Kerscher A and Slack FJ: Oncomirs-microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar : PubMed/NCBI
25	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
26	Kriegel AJ, Liu Y, Fang Y, Ding X and Liang M: The miR-29 family: Genomics, cell biology, and relevance to renal and cardiovascular injury. Physiol Genomics. 44:237–244. 2012. View Article : Google Scholar : PubMed/NCBI
27	Wang Y, Zhang X, Li H, Yu J and Ren X: The role of miRNA-29 family in cancer. Eur J Cell Biol. 92:123–128. 2013. View Article : Google Scholar : PubMed/NCBI
28	Li J, Wan X, Qiang W, Li T, Huang W, Huang S, Wu D and Li Y: MiR-29a suppresses prostate cell proliferation and induces apoptosis via KDM5B protein regulation. Int J Clin Exp Med. 8:5329–5339. 2015.PubMed/NCBI
29	Liu X, Cai J, Sun Y, Gong R, Sun D, Zhong X, Jiang S, He X, Bao E, Yang L and Li Y: MicroRNA-29a inhibits cell migration and invasion via targeting Roundabout homolog 1 in gastric cancer cells. Mol Med Rep. 12:3944–3950. 2015.PubMed/NCBI
30	Liu C, Duan P, Li B, Huang C, Jing Y and Yan W: miR-29a activates Hes1 by targeting Nfia in esophageal carcinoma cell line TE-1. Oncol Lett. 9:96–102. 2015.PubMed/NCBI
31	Wang F, Wang XS, Yang GH, Zhai PF, Xiao Z, Xia LY, Chen LR, Wang Y, Wang XZ, Bi LX, et al: miR-29a and miR-142-3p downregulation and diagnostic implication in human acute myeloid leukemia. Mol Biol Rep. 39:2713–2722. 2012. View Article : Google Scholar : PubMed/NCBI
32	Zhu XC, Dong QZ, Zhang XF, Deng B, Jia HL, Ye QH, Qin LX and Wu XZ: microRNA-29a suppresses cell proliferation by targeting SPARC in hepatocellular carcinoma. Int J Mol Med. 30:1321–1326. 2012.PubMed/NCBI
33	Tang W, Zhu Y, Gao J, Fu J, Liu C, Liu Y, Song C, Zhu S, Leng Y, Wang G, et al: MicroRNA-29a promotes colorectal cancer metastasis by regulating matrix metalloproteinase 2 and E-cadherin via KLF4. Br J Cancer. 110:450–458. 2014. View Article : Google Scholar : PubMed/NCBI
34	Nishikawa R, Chiyomaru T, Enokida H, Inoguchi S, Ishihara T, Matsushita R, Goto Y, Fukumoto I, Nakagawa M and Seki N: Tumour-suppressive microRNA-29s directly regulate LOXL2 expression and inhibit cancer cell migration and invasion in renal cell carcinoma. FEBS Lett. 589:2136–2145. 2015. View Article : Google Scholar : PubMed/NCBI
35	Tréhoux S, Lahdaoui F, Delpu Y, Renaud F, Leteurtre E, Torrisani J, Jonckheere N and Van Seuningen I: Micro-RNAs miR-29a and miR-330-5p function as tumor suppressors by targeting the MUC1 mucin in pancreatic cancer cells. Biochim Biophys Acta. 1853:2392–2403. 2015. View Article : Google Scholar : PubMed/NCBI
36	Zhao Z, Wang L, Song W, Cui H, Chen G, Qiao F, Hu J, Zhou R and Fan H: Reduced miR-29a-3p expression is linked to the cell proliferation and cell migration in gastric cancer. World J Surg Oncol. 13:1012015. View Article : Google Scholar : PubMed/NCBI
37	Lu L, Xue X, Lan J, Gao Y, Xiong Z, Zhang H, Jiang W, Song W and Zhi Q: MicroRNA-29a upregulates MMP2 in oral squamous cell carcinoma to promote cancer invasion and anti-apoptosis. Biomed Pharmacother. 68:13–19. 2014. View Article : Google Scholar : PubMed/NCBI
38	Ma J, Xue Y, Liu W, Yue C, Bi F, Xu J, Zhang J, Li Y, Zhong C and Chen Y: Role of activated Rac1/Cdc42 in mediating endothelial cell proliferation and tumor angiogenesis in breast cancer. PLoS One. 8:e662752013. View Article : Google Scholar : PubMed/NCBI
39	Sun N, Ye L, Chang T and Li X and Li X: microRNA-195-Cdc42 axis acts as a prognostic factor of esophageal squamous cell carcinoma. Int J Clin Exp Pathol. 7:6871–6879. 2014.PubMed/NCBI
40	Reymond N, Im JH, Garg R, Vega FM, Borda d'Agua B, Riou P, Cox S, Valderrama F, Muschel RJ and Ridley AJ: Cdc42 promotes transendothelial migration of cancer cells through β1 integrin. J Cell Biol. 199:653–668. 2012. View Article : Google Scholar : PubMed/NCBI
41	Stengel KR and Zheng Y: Essential role of Cdc42 in Ras-induced transformation revealed by gene targeting. PLoS One. 7:e373172012. View Article : Google Scholar : PubMed/NCBI
42	Nobes CD and Hall A: Rho, rac and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell. 81:53–62. 1995. View Article : Google Scholar : PubMed/NCBI
43	Olson MF, Ashworth A and Hall A: An essential role for Rho, Rac and Cdc42 GTPases in cell cycle progression through G1. Science. 269:1270–1272. 1995. View Article : Google Scholar : PubMed/NCBI
44	Chen QY, Jiao DM, Yao QH, Yan J, Song J, Chen FY, Lu GH and Zhou JY: Expression analysis of Cdc42 in lung cancer and modulation of its expression by curcumin in lung cancer cell lines. Int J Oncol. 40:1561–1568. 2012.PubMed/NCBI
45	Liu M, Lang N, Qiu M, Xu F, Li Q, Tang Q, Chen J, Chen X, Zhang S, Liu Z, et al: miR-137 targets Cdc42 expression, induces cell cycle G1 arrest and inhibits invasion in colorectal cancer cells. Int J Cancer. 128:1269–1279. 2011. View Article : Google Scholar : PubMed/NCBI
46	Humphreys KJ, McKinnon RA and Michael MZ: miR-18a inhibits CDC42 and plays a tumour suppressor role in colorectal cancer cells. PLoS One. 9:e1122882014. View Article : Google Scholar : PubMed/NCBI
47	Ke TW, Hsu HL, Wu YH, Chen WT, Cheng YW and Cheng CW: MicroRNA-224 suppresses colorectal cancer cell migration by targeting Cdc42. Dis Markers. 2014:6171502014. View Article : Google Scholar : PubMed/NCBI
48	Fu MG, Li S, Yu TT, Qian LJ, Cao RS, Zhu H, Xiao B, Jiao CH, Tang NN, Ma JJ, et al: Differential expression of miR-195 in esophageal squamous cell carcinoma and miR-195 expression inhibits tumor cell proliferation and invasion by targeting of Cdc42. FEBS Lett. 587:3471–3479. 2013. View Article : Google Scholar : PubMed/NCBI
49	Zhang Y, Takahashi S, Tasaka A, Yoshima T, Ochi H and Chayama K: Involvement of microRNA-224 in cell proliferation, migration, invasion, and anti-apoptosis in hepatocellular carcinoma. J Gastroenterol Hepatol. 28:565–575. 2013. View Article : Google Scholar : PubMed/NCBI
50	Chen Q, Chen X, Zhang M, Fan Q, Luo S and Cao X: miR-137 is frequently down-regulated in gastric cancer and is a negative regulator of Cdc42. Dig Dis Sci. 56:2009–2016. 2011. View Article : Google Scholar : PubMed/NCBI
51	Yang T, Chen T, Li Y, Gao L, Zhang S, Wang T and Chen M: Downregulation of miR-25 modulates non-small cell lung cancer cells by targeting CDC42. Tumour Biol. 36:1903–1911. 2015. View Article : Google Scholar : PubMed/NCBI
52	Zhu X, Li Y, Shen H, Li H, Long L, Hui L and Xu W: miR-137 inhibits the proliferation of lung cancer cells by targeting Cdc42 and Cdk6. FEBS Lett. 587:73–81. 2013. View Article : Google Scholar : PubMed/NCBI