Downregulation of microRNA‑1246 inhibits tumor growth and promotes apoptosis of cervical cancer cells by targeting thrombospondin‑2

Du,Ping; Lai,Yue‑Hua; Yao,De‑Sheng; Chen,Jun‑Ying; Ding,Nan

doi:10.3892/ol.2019.10571

Downregulation of microRNA‑1246 inhibits tumor growth and promotes apoptosis of cervical cancer cells by targeting thrombospondin‑2

Authors:
- Ping Du
- Yue‑Hua Lai
- De‑Sheng Yao
- Jun‑Ying Chen
- Nan Ding
View Affiliations

Affiliations: Department of Gynecological Oncology, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Gynecology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530022, P.R. China
Published online on: July 5, 2019 https://doi.org/10.3892/ol.2019.10571
Pages: 2491-2499
Copyright: © Du 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

Cervical cancer pathogenesis is regulated by numerous factors, including microRNAs. MicroRNA 1246 (miR‑1246) has been shown to serve a role in cervical cancer tumorigenesis. However, the mechanisms through which miR‑1246 exerts its oncogenic effects are largely unknown. The aim of the current study was to evaluate the effects of lentivirus‑mediated miR‑1246 knockdown on the biological characteristics and behavior of cervical cancer cells, and to identify the downstream signaling pathways affected by miR‑1246 knockdown. Short hairpins inhibiting miR‑1246 were synthesized and cloned into a recombinant lentiviral vector (LV‑miR‑1246‑Inh), which was then used to infect SiHa cervical cancer cells. The effects of LV‑miR‑1246‑Inh infection on cell invasion, proliferation and apoptosis were evaluated by Transwell assay, Cell Counting Kit‑8 assay and flow cytometry, respectively. Thrombospondin‑2 (THBS2), matrix metalloproteinase 2 (MMP2), MMP9 and extracellular matrix (ECM) component expression levels were evaluated, and the growth of xenograft tumors formed following injection of SiHa cells with knockdown of miR‑1246 was assessed. miR‑1246 downregulation in SiHa cells decreased proliferation, induced apoptosis and upregulated THBS2 expression. Furthermore, MMP2 and MMP9 levels were downregulated, whereas components of the ECM were upregulated subsequent to miR‑1246 knockdown, indicating that this miRNA regulates cervical cancer cell pathogenesis via the THBS2/MMP/ECM pathway. Notably, SiHa cells with miR‑1246 downregulation had a markedly decreased ability to form tumors in vivo. These results suggest that miR‑1246 functions during cervical cancer pathogenesis and tumor formation via the THBS2/MMP/ECM signaling pathway. These findings support the future use of miR‑1246 suppression in the treatment of cervical cancer.

View Figures

View References

1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	Cibula D, Abu-Rustum NR, Fischerova D, Pather S, Lavigne K, Slama J, Alektiar K, Ming-Yin L, Kocian R, Germanova A, et al: Surgical treatment of ‘intermediate risk’ lymph node negative cervical cancer patients without adjuvant radiotherapy-A retrospective cohort study and review of the literature. Gynecol Oncol. 151:438–443. 2018. View Article : Google Scholar : PubMed/NCBI
3	Schwarz DS, Hutvagner G, Du T, Xu ZS, Aronin N and Zamore PD: Asymmetry in the assembly of the RNAi enzyme complex. Cell. 115:199–208. 2003. View Article : Google Scholar : PubMed/NCBI
4	Gregory RI, Chendrimada TP, Cooch N and Shiekhattar R: Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. Cell. 123:631–640. 2005. View Article : Google Scholar : PubMed/NCBI
5	Pedroza-Torres A, Lopez-Urrutia E, Garcia-Castillo V, Jacobo-Herrera N, Herrera LA, Peralta-Zaragoza O, López-Camarillo C, De Leon DC, Fernández-Retana J, Cerna-Cortés JF, et al: MicroRNAs in cervical cancer: Evidences for a miRNA profile deregulated by HPV and its impact on radio-resistance. Molecules. 19:6263–6281. 2014. View Article : Google Scholar : PubMed/NCBI
6	Hu X, Schwarz JK, Lewis JS Jr, Huettner PC, Rader JS, Deasy JO, Grigsby PW and Wang X: A microRNA expression signature for cervical cancer prognosis. Cancer Res. 70:1441–1448. 2010. View Article : Google Scholar : PubMed/NCBI
7	Ribeiro J and Sousa H: MicroRNAs as biomarkers of cervical cancer development: A literature review on miR-125b and miR-34a. Mol Biol Rep. 41:1525–1531. 2014. View Article : Google Scholar : PubMed/NCBI
8	Fiannaca A, La Rosa M, La Paglia L, Rizzo R and Urso A: Analysis of miRNA expression profiles in breast cancer using biclustering. BMC Bioinformatics. 16 (Suppl 4):S72015. View Article : Google Scholar : PubMed/NCBI
9	Liao JM, Zhou X, Zhang Y and Lu H: MiR-1246: A new link of the p53 family with cancer and Down syndrome. Cell Cycle. 11:2624–2630. 2012. View Article : Google Scholar : PubMed/NCBI
10	Chen J, Yao D, Li Y, Chen H, He C, Ding N, Lu Y, Ou T, Zhao S, Li L and Long F: Serum microRNA expression levels can predict lymph node metastasis in patients with early-stage cervical squamous cell carcinoma. Int J Mol Med. 32:557–567. 2013. View Article : Google Scholar : PubMed/NCBI
11	Chen J, Yao D, Zhao S, He C, Ding N, Li L and Long F: MiR-1246 promotes SiHa cervical cancer cell proliferation, invasion, and migration through suppression of its target gene thrombospondin 2. Arch Gynecol Obstet. 290:725–732. 2014. View Article : Google Scholar : PubMed/NCBI
12	Bao Y, Wang L, Shi L, Yun F, Liu X, Chen Y, Chen C, Ren Y and Jia Y: Transcriptome profiling revealed multiple genes and ECM-receptor interaction pathways that may be associated with breast cancer. Cell Mol Biol Lett. 24:382019. View Article : Google Scholar : PubMed/NCBI
13	Yang Z, Kyriakides TR and Bornstein P: Matricellular proteins as modulators of cell-matrix interactions: Adhesive defect in thrombospondin 2-null fibroblasts is a consequence of increased levels of matrix metalloproteinase-2. Mol Biol Cell. 11:3353–3364. 2000. View Article : Google Scholar : PubMed/NCBI
14	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
15	Naito S, von Eschenbach AC, Giavazzi R and Fidler IJ: Growth and metastasis of tumor cells isolated from a human renal cell carcinoma implanted into different organs of nude mice. Cancer Res. 46:4109–4115. 1986.PubMed/NCBI
16	Menon SS, Guruvayoorappan C, Sakthivel KM and Rasmi RR: Ki-67 protein as a tumour proliferation marker. Clin Chim Acta. 491:39–45. 2019. View Article : Google Scholar : PubMed/NCBI
17	Varghese F, Bukhari AB, Malhotra R and De A: IHC Profiler: An open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples. PLoS One. 9:e968012014. View Article : Google Scholar : PubMed/NCBI
18	Weidner N, Semple JP, Welch WR and Folkman J: Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med. 324:1–8. 1991. View Article : Google Scholar : PubMed/NCBI
19	He L, He X, Lowe SW and Hannon GJ: microRNAs join the p53 network-another piece in the tumour-suppression puzzle. Nat Rev Cancer. 7:819–822. 2007. View Article : Google Scholar : PubMed/NCBI
20	Yang Y, Liu Y, Li G, Li L, Geng P and Song H: microRNA-214 suppresses the growth of cervical cancer cells by targeting EZH2. Oncol Lett. 16:5679–5686. 2018.PubMed/NCBI
21	Pedroza-Torres A, Campos-Parra AD, Millan-Catalan O, Loissell-Baltazar YA, Zamudio-Meza H, Cantú de León D, Montalvo-Esquivel G, Isla-Ortiz D, Herrera LA, Ángeles-Zaragoza Ó, et al: MicroRNA-125 modulates radioresistance through targeting p21 in cervical cancer. Oncol Rep. 39:1532–1540. 2018.PubMed/NCBI
22	González-Quintana V, Palma-Berré L, Campos-Parra AD, López-Urrutia E, Peralta-Zaragoza O, Vazquez-Romo R and Pérez-Plasencia C: MicroRNAs are involved in cervical cancer development, progression, clinical outcome and improvement treatment response (Review). Oncol Rep. 35:3–12. 2016. View Article : Google Scholar : PubMed/NCBI
23	Morin RD, O'Connor MD, Griffith M, Kuchenbauer F, Delaney A, Prabhu AL, Zhao Y, McDonald H, Zeng T, Hirst M, et al: Application of massively parallel sequencing to microRNA profiling and discovery in human embryonic stem cells. Genome Res. 18:610–621. 2008. View Article : Google Scholar : PubMed/NCBI
24	Krissansen GW, Yang Y, McQueen FM, Leung E, Peek D, Chan YC, Print C, Dalbeth N, Williams M and Fraser AG: Overexpression of miR-595 and miR-1246 in the sera of patients with active forms of inflammatory bowel disease. Inflamm Bowel Dis. 21:520–530. 2015. View Article : Google Scholar : PubMed/NCBI
25	Todeschini P, Salviato E, Paracchini L, Ferracin M, Petrillo M, Zanotti L, Tognon G, Gambino A, Calura E, Caratti G, et al: Circulating miRNA landscape identifies miR-1246 as promising diagnostic biomarker in high-grade serous ovarian carcinoma: A validation across two independent cohorts. Cancer Lett. 388:320–327. 2017. View Article : Google Scholar : PubMed/NCBI
26	Zhang WC, Chin TM, Yang H, Nga ME, Lunny DP, Lim EK, Sun LL, Pang YH, Leow YN, Malusay SR, et al: Tumour-initiating cell-specific miR-1246 and miR-1290 expression converge to promote non-small cell lung cancer progression. Nat Commun. 7:117022016. View Article : Google Scholar : PubMed/NCBI
27	Kim G, An HJ, Lee MJ, Song JY, Jeong JY, Lee JH and Jeong HC: Hsa-miR-1246 and hsa-miR-1290 are associated with stemness and invasiveness of non-small cell lung cancer. Lung Cancer. 91:15–22. 2016. View Article : Google Scholar : PubMed/NCBI
28	Hasegawa S, Eguchi H, Nagano H, Konno M, Tomimaru Y, Wada H, Hama N, Kawamoto K, Kobayashi S, Nishida N, et al: MicroRNA-1246 expression associated with CCNG2-mediated chemoresistance and stemness in pancreatic cancer. Br J Cancer. 111:1572–1580. 2014. View Article : Google Scholar : PubMed/NCBI
29	Takeshita N, Hoshino I, Mori M, Akutsu Y, Hanari N, Yoneyama Y, Ikeda N, Isozaki Y, Maruyama T, Akanuma N, et al: Serum microRNA expression profile: miR-1246 as a novel diagnostic and prognostic biomarker for oesophageal squamous cell carcinoma. Br J Cancer. 108:644–652. 2013. View Article : Google Scholar : PubMed/NCBI
30	Kishi M, Nakamura M, Nishimine M, Ishida E, Shimada K, Kirita T and Konishi N: Loss of heterozygosity on chromosome 6q correlates with decreased thrombospondin-2 expression in human salivary gland carcinomas. Cancer Sci. 94:530–535. 2003. View Article : Google Scholar : PubMed/NCBI
31	Ao R, Guan L, Wang Y and Wang JN: Silencing of COL1A2, COL6A3, and THBS2 inhibits gastric cancer cell proliferation, migration, and invasion while promoting apoptosis through the PI3k-Akt signaling pathway. J Cell Biochem. 119:4420–4434. 2018. View Article : Google Scholar : PubMed/NCBI
32	Tokunaga T, Nakamura M, Oshika Y, Abe Y, Ozeki Y, Fukushima Y, Hatanaka H, Sadahiro S, Kijima H, Tsuchida T, et al: Thrombospondin 2 expression is correlated with inhibition of angiogenesis and metastasis of colon cancer. Br J Cancer. 79:354–359. 1999. View Article : Google Scholar : PubMed/NCBI
33	Weng TY, Wang CY, Hung YH, Chen WC, Chen YL and Lai MD: Differential expression pattern of THBS1 and THBS2 in lung cancer: Clinical outcome and a systematic-analysis of microarray databases. PLoS One. 11:e1610072016. View Article : Google Scholar
34	Fears CY, Grammer JR, Stewart JE Jr, Annis DS, Mosher DF, Bornstein P and Gladson CL: Low-density lipoprotein receptor-related protein contributes to the antiangiogenic activity of thrombospondin-2 in a murine glioma model. Cancer Res. 65:9338–9346. 2005. View Article : Google Scholar : PubMed/NCBI
35	Nakamura M, Oida Y, Abe Y, Yamazaki H, Mukai M, Matsuyama M, Chijiwa T, Matsumoto H and Ueyama Y: Thrombospondin-2 inhibits tumor cell invasion through the modulation of MMP-9 and uPA in pancreatic cancer cells. Mol Med Rep. 1:423–427. 2008.PubMed/NCBI
36	Kodama J, Hashimoto I, Seki N, Hongo A, Yoshinouchi M, Okuda H and Kudo T: Thrombospondin-1 and −2 messenger RNA expression in invasive cervical cancer: Correlation with angiogenesis and prognosis. Clin Cancer Res. 7:2826–2831. 2001.PubMed/NCBI
37	Kamochi J, Tokunaga T, Tomii Y, Abe Y, Hatanaka H, Kijima H, Yamazaki H, Watanabe N, Matsuzaki S, Ueyama Y and Nakamura M: Overexpression of the thrombospondin 2 (TSP2) gene modulated by the matrix metalloproteinase family expression and production in human colon carcinoma cell line. Oncol Rep. 10:881–884. 2003.PubMed/NCBI
38	Chen PC, Tang CH, Lin LW, Tsai CH, Chu CY, Lin TH and Huang YL: Thrombospondin-2 promotes prostate cancer bone metastasis by the up-regulation of matrix metalloproteinase-2 through down-regulating miR-376c expression. J Hematol Oncol. 10:332017. View Article : Google Scholar : PubMed/NCBI
39	Davies KJ: The complex interaction of matrix metalloproteinases in the migration of cancer cells through breast tissue stroma. Int J Breast Cancer. 2014:8390942014. View Article : Google Scholar : PubMed/NCBI
40	Hirose Y, Chiba K, Karasugi T, Nakajima M, Kawaguchi Y, Mikami Y, Furuichi T, Mio F, Miyake A, Miyamoto T, et al: A functional polymorphism in THBS2 that affects alternative splicing and MMP binding is associated with lumbar-disc herniation. Am J Hum Genet. 82:1122–1129. 2008. View Article : Google Scholar : PubMed/NCBI