lnc-PKD2-2-3, identified by long non-coding RNA expression profiling, is associated with pejorative tumor features and poor prognosis, enhances cancer stemness and may serve as cancer stem-cell marker in cholangiocarcinoma

Qiu,Gongcai; Ma,Donglai; Li,Fujun; Sun,Dongsheng; Zeng,Zhaolin

doi:10.3892/ijo.2019.4798

lnc-PKD2-2-3, identified by long non-coding RNA expression profiling, is associated with pejorative tumor features and poor prognosis, enhances cancer stemness and may serve as cancer stem-cell marker in cholangiocarcinoma

Authors:
- Gongcai Qiu
- Donglai Ma
- Fujun Li
- Dongsheng Sun
- Zhaolin Zeng
View Affiliations

Affiliations: Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: May 6, 2019 https://doi.org/10.3892/ijo.2019.4798
Pages: 45-58
Copyright: © Qiu 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

The present study aimed to explore the long non‑coding RNA (lncRNA) expression profiles and correlation of lnc‑PKD2‑2‑3 with tumor features and prognosis, and to investigate its effect on regulating cancer‑cell stemness and its potential as a cancer stem cell (CSC) marker in cholangiocarcinoma (CCA). lncRNA expression profiles were determined in 3 pairs of CCA tumors and adjacent tissues by microarray analysis, and lnc‑PKD2‑2‑3 expression was then validated in 60 paired samples by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Expression of common CSC markers [(CD44, CD133 and octamer‑binding transcription factor 4 (OCT4)], CD44+CD133+ cell proportions, sphere formation efficiency and drug resistance to 5‑fluorouracil (5‑FU) were measured following ectopic overexpression of lnc‑PKD2‑2‑3 or silencing via small hairpin RNA lentivirus transfection into the TFK‑1 and Huh‑28 CCA cell lines. Finally, lnc‑PKD2‑2‑3 expression was measured in CCA stem‑like cells and normal CCA cells. The results from the microarray analysis identified a total of 4,223 upregulated and 4,596 downregulated lncRNAs between CCA tumor tissue and paired adjacent tissue, which were enriched in regulating cancer‑associated pathways. RT‑qPCR validation revealed that lnc‑PKD2‑2‑3 was upregulated in CCA and associated with a higher Eastern Cooperative Oncology Group performance score, poor differentiation, advanced TNM stage, increased carcinoembryonic antigen and poor overall survival in CCA patients. In vitro, lnc‑PKD2‑2‑3 increased CD44, CD133 and OCT4 expression as well as the CD44+CD133+ cell proportion, raised the sphere formation efficiency and enhanced drug resistance to 5‑FU in TFK‑1 and Huh‑28 cells. In addition, lnc‑PKD2‑2‑3 was positively correlated with CSC markers in CCA tumor tissues and was markedly upregulated in CCA stem‑like cells compared with that in normal CCA cells. In conclusion, lnc‑PKD2‑2‑3, selected by lncRNA expression profiling, was associated with pejorative tumor features and poor prognosis, enhanced cancer stemness and may serve as a CSC marker in CCA.

View Figures

View References

1	Shin HR, Oh JK, Masuyer E, Curado MP, Bouvard V, Fang YY, Wiangnon S, Sripa B and Hong ST: Epidemiology of cholangiocarcinoma: An update focusing on risk factors. Cancer Sci. 101:579–585. 2010. View Article : Google Scholar : PubMed/NCBI
2	Mosadeghi S, Liu B, Bhuket T and Wong RJ: Sex-specific and race/ethnicity-specific disparities in cholangiocarcinoma incidence and prevalence in the USA: An updated analysis of the 2000-2011 Surveillance, Epidemiology and End Results registry. Hepatol Res. 46:669–677. 2016. View Article : Google Scholar
3	Razumilava N and Gores GJ: Cholangiocarcinoma. Lancet. 383:2168–2179. 2014. View Article : Google Scholar : PubMed/NCBI
4	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
5	Rizvi S, Khan SA, Hallemeier CL, Kelley RK and Gores GJ: Cholangiocarcinoma - evolving concepts and therapeutic strategies. Nat Rev Clin Oncol. 15:95–111. 2018. View Article : Google Scholar
6	Bridgewater J, Galle PR, Khan SA, Llovet JM, Park JW, Patel T, Pawlik TM and Gores GJ: Guidelines for the diagnosis and management of intrahepatic cholangiocarcinoma. J Hepatol. 60:1268–1289. 2014. View Article : Google Scholar : PubMed/NCBI
7	Rizvi S and Gores GJ: Emerging molecular therapeutic targets for cholangiocarcinoma. J Hepatol. 67:632–644. 2017. View Article : Google Scholar : PubMed/NCBI
8	Balas MM and Johnson AM: Exploring the mechanisms behind long noncoding RNAs and cancer. Noncoding RNA Res. 3:108–117. 2018. View Article : Google Scholar : PubMed/NCBI
9	Rafiee A, Riazi-Rad F, Havaskary M and Nuri F: Long noncoding RNAs: Regulation, function and cancer. Biotechnol Genet Eng Rev. 34:153–180. 2018. View Article : Google Scholar : PubMed/NCBI
10	Evans JR, Feng FY and Chinnaiyan AM: The bright side of dark matter: lncRNAs in cancer. J Clin Invest. 126:2775–2782. 2016. View Article : Google Scholar : PubMed/NCBI
11	Schmitt AM and Chang HY: Long noncoding RNAs in cancer pathways. Cancer Cell. 29:452–463. 2016. View Article : Google Scholar : PubMed/NCBI
12	Prensner JR and Chinnaiyan AM: The emergence of lncRNAs in cancer biology. Cancer Discov. 1:391–407. 2011. View Article : Google Scholar : PubMed/NCBI
13	Song H, Xu Y, Shi L, Xu T, Fan R, Cao M, Xu W and Song J: LncRNA THOR increases the stemness of gastric cancer cells via enhancing SOX9 mRNA stability. Biomed Pharmacother. 108:338–346. 2018. View Article : Google Scholar : PubMed/NCBI
14	Wang Y, He L, Du Y, Zhu P, Huang G, Luo J, Yan X, Ye B, Li C, Xia P, et al: The long noncoding RNA lncTCF7 promotes self-renewal of human liver cancer stem cells through activation of Wnt signaling. Cell Stem Cell. 16:413–425. 2015. View Article : Google Scholar : PubMed/NCBI
15	Zhang Z, Sun L, Zhang Y, Lu G, Li Y and Wei Z: Long non-coding RNA FEZF1-AS1 promotes breast cancer stemness and tumorigenesis via targeting miR-30a/Nanog axis. J Cell Physiol. 233:8630–8638. 2018. View Article : Google Scholar : PubMed/NCBI
16	Batlle E and Clevers H: Cancer stem cells revisited. Nat Med. 23:1124–1134. 2017. View Article : Google Scholar : PubMed/NCBI
17	Pützer BM, Solanki M and Herchenröder O: Advances in cancer stem cell targeting: How to strike the evil at its root. Adv Drug Deliv Rev. 120:89–107. 2017. View Article : Google Scholar : PubMed/NCBI
18	Li Y, Cai Q, Li W, Feng F and Yang L: Long non-coding RNA EPIC1 promotes cholangiocarcinoma cell growth. Biochem Biophys Res Commun. 504:654–659. 2018. View Article : Google Scholar : PubMed/NCBI
19	Guo L, Zhou Y, Chen Y, Sun H, Wang Y and Qu Y: LncRNA ASAP1-IT1 positively modulates the development of cholangiocarcinoma via hedgehog signaling pathway. Biomed Pharmacother. 103:167–173. 2018. View Article : Google Scholar : PubMed/NCBI
20	Li Z, Li J, Ji D, Leng K, Xu Y, Huang L, Jiang X and Cui Y: Overexpressed long noncoding RNA Sox2ot predicts poor prognosis for cholangiocarcinoma and promotes cell proliferation and invasion. Gene. 645:131–136. 2018. View Article : Google Scholar
21	Huang W, Sherman BT and Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 4:44–57. 2009. View Article : Google Scholar
22	Huang W, Sherman BT and Lempicki RA: Bioinformatics enrichment tools: Paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 37:1–13. 2009. View Article : Google Scholar :
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
24	Kumar S, Williams D, Sur S, Wang JY and Jo H: Role of flow-sensitive microRNAs and long noncoding RNAs in vascular dysfunction and atherosclerosis. Vascul Pharmacol. 114:76–92. 2019. View Article : Google Scholar
25	Feng SD, Yang JH, Yao CH, Yang SS, Zhu ZM, Wu D, Ling HY and Zhang L: Potential regulatory mechanisms of lncRNA in diabetes and its complications. Biochem Cell Biol. 95:361–367. 2017. View Article : Google Scholar : PubMed/NCBI
26	Quan J, Pan X, Zhao L, Li Z, Dai K, Yan F, Liu S, Ma H and Lai Y: LncRNA as a diagnostic and prognostic biomarker in bladder cancer: A systematic review and meta-analysis. OncoTargets Ther. 11:6415–6424. 2018. View Article : Google Scholar
27	Wang X, Liu F, Liu X, Wang F, Liao X, Chen Y, Mao Y, Hua D and Ge X: Long non-coding RNA expression profiles reveals AK098783 is a biomarker to predict poor prognosis in patients with colorectal cancer. Jpn J Clin Oncol. 48:480–484. 2018. View Article : Google Scholar : PubMed/NCBI
28	Yao J, Wu L, Meng X, Yang H, Ni S, Wang Q, Zhou J, Zhang Q, Su K, Shao L, et al: Profiling, clinicopathological correlation and functional validation of specific long non-coding RNAs for hepatocellular carcinoma. Mol Cancer. 16:1642017. View Article : Google Scholar : PubMed/NCBI
29	Ma J, Zhang L, Yang P, Zeng ZC and Xiang ZL: Integrated analysis of long noncoding RNA expression profiles in lymph node metastasis of hepatocellular carcinoma. Gene. 676:47–55. 2018. View Article : Google Scholar : PubMed/NCBI
30	Zhang F, Wan M, Xu Y, Li Z, Kang P, Jiang X, Wang Y, Wang Z, Zhong X, Li C, et al: Transcriptome analysis reveals dysregulated long non-coding RNAs and mRNAs associated with extrahepatic cholangiocarcinoma progression. Oncol Lett. 14:6079–6084. 2017.PubMed/NCBI
31	Zhang W, Zhai Y, Wang W, Cao M and Ma C: Enhanced expression of lncRNA TP73-AS1 predicts unfavorable prognosis for gastric cancer and promotes cell migration and invasion by induction of EMT. Gene. 678:377–383. 2018. View Article : Google Scholar : PubMed/NCBI
32	Li C, Li W, Zhang Y, Zhang X, Liu T, Zhang Y, Yang Y, Wang L, Pan H, Ji J, et al: Increased expression of antisense lncRNA SPINT1-AS1 predicts a poor prognosis in colorectal cancer and is negatively correlated with its sense transcript. OncoTargets Ther. 11:3969–3978. 2018. View Article : Google Scholar
33	Jin L, He Y, Tang S and Huang S: LncRNA GHET1 predicts poor prognosis in hepatocellular carcinoma and promotes cell proliferation by silencing KLF2. J Cell Physiol. 233:4726–4734. 2018. View Article : Google Scholar
34	Carnero A, Garcia-Mayea Y, Mir C, Lorente J, Rubio IT and LLeonart ME: The cancer stem-cell signaling network and resistance to therapy. Cancer Treat Rev. 49:25–36. 2016. View Article : Google Scholar : PubMed/NCBI
35	Ravasio R, Ceccacci E and Minucci S: Self-renewal of tumor cells: Epigenetic determinants of the cancer stem cell phenotype. Curr Opin Genet Dev. 36:92–99. 2016. View Article : Google Scholar : PubMed/NCBI
36	Cardinale V, Renzi A, Carpino G, Torrice A, Bragazzi MC, Giuliante F, DeRose AM, Fraveto A, Onori P, Napoletano C, et al: Profiles of cancer stem cell subpopulations in cholangiocarcinomas. Am J Pathol. 185:1724–1739. 2015. View Article : Google Scholar : PubMed/NCBI
37	Kemmerling R, Alinger B, Dietze O, Bösmüller HC, Ocker M, Wolkersdörfer GW, Berr F, Neureiter D and Kiesslich T: Association of stem cell marker expression pattern and survival in human biliary tract cancer. Int J Oncol. 41:511–522. 2012. View Article : Google Scholar : PubMed/NCBI
38	Yoshikawa S, Zen Y, Fujii T, Sato Y, Ohta T, Aoyagi Y and Nakanuma Y: Characterization of CD133⁺ parenchymal cells in the liver: Histology and culture. World J Gastroenterol. 15:4896–4906. 2009. View Article : Google Scholar : PubMed/NCBI