Single‑cell intratumoral stemness analysis reveals the involvement of cell cycle and DNA damage repair in two different types of esophageal cancer

Wu,Hongjin; Li,Ying; Hou,Qiang; Zhou,Rongjin; Li,Ziwei; Wu,Shixiu; Yu,Juehua; Jiang,Mingfeng

doi:10.3892/or.2019.7117

Single‑cell intratumoral stemness analysis reveals the involvement of cell cycle and DNA damage repair in two different types of esophageal cancer

Authors:
- Hongjin Wu
- Ying Li
- Qiang Hou
- Rongjin Zhou
- Ziwei Li
- Shixiu Wu
- Juehua Yu
- Mingfeng Jiang
View Affiliations

Affiliations: Cancer Research Institute, Hangzhou Cancer Hospital, Hangzhou, Zhejiang 320000, P.R. China, Yunnan Institute of Digestive Disease, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650032, P.R. China
Published online on: April 15, 2019 https://doi.org/10.3892/or.2019.7117
Pages: 3201-3208
Copyright: © Wu 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

Intratumoral heterogeneity, particularly the potential cancer stemness of single cancer cells, has not yet been fully elucidated in human esophageal cancer. Single‑cell transcriptome sequencing of two types of esophageal adenocarcinoma (EAC) and two types of esophageal squamous cell carcinoma (ESCC) tissues was performed, and the intratumoral cancer stemness of the types of esophageal cancer were characterized at the single‑cell level in the present study. By comparing the transcriptomic profiles of single cancer cells with high and low stemness in individual patients, it was revealed that the overexpression of cell cycle‑associated genes in EAC cells was highly correlated with stemness, whereas overexpression of genes involved in the signaling pathways of DNA replication and DNA damage repair was significantly correlated with stemness in ESCC. High expression of these stemness‑associated genes was correlated with poor prognosis of patients. Additionally, poly [ADP‑ribose] polymerase(PARP)4 was identified as a novel cancer stemness‑associated gene in ESCC and its association with survival was validated in a cohort of 121 patients with ESCC. These findings have profound potential implications for the use of cell cycle inhibitors in EAC and PARP inhibitors in ESCC, which may provide novel mechanistic insights into the plasticity of esophageal cancer.

View Figures

View References

1	Wong DJ, Segal E and Chang HY: Stemness, cancer and cancer stem cells. Cell Cycle. 7:3622–3624. 2008. View Article : Google Scholar : PubMed/NCBI
2	Monteiro J and Fodde R: Cancer stemness and metastasis: Therapeutic consequences and perspectives. Eur J Cancer. 46:1198–1203. 2010. View Article : Google Scholar : PubMed/NCBI
3	Khosrotehrani K and Roy E: Tell me about your stemness. I'll give your cancer risk! Cell Death Differ. 24:6–7. 2017. View Article : Google Scholar
4	Ghisolfi L, Keates AC, Hu X, Lee DK and Li CJ: Ionizing radiation induces stemness in cancer cells. PLoS One. 7:e436282012. View Article : Google Scholar : PubMed/NCBI
5	Hu X, Ghisolfi L, Keates AC, Zhang J, Xiang S and Lee DK: Induction of cancer cell stemness by chemotherapy. Cell Cycle. 11:2691–2698. 2012. View Article : Google Scholar : PubMed/NCBI
6	Zhang Z, Duan Q, Zhao H, Liu T, Wu H, Shen Q, Wang C and Yin T: Gemcitabine treatment promotes pancreatic cancer stemness through the Nox/ROS/NF-kappaB/STAT3 signaling cascade. Cancer Lett. 382:53–63. 2016. View Article : Google Scholar : PubMed/NCBI
7	Liu L, Yang L, Yan W, Zhai J, Pizzo DP, Chu P, Chin AR, Shen M, Dong C, Ruan X, et al: Chemotherapy induces breast cancer stemness in association with dysregulated monocytosis. Clin Cancer Res. 24:2370–2382. 2018. View Article : Google Scholar : PubMed/NCBI
8	Li Y, Rogoff HA, Keates S, Gao Y, Murikipudi S, Mikule K, Leggett D, Li W, Pardee AB and Li CJ: Suppression of cancer relapse and metastasis by inhibiting cancer stemness. Proc Natl Acad Sci USA. 112:1839–1844. 2015. View Article : Google Scholar : PubMed/NCBI
9	Liang ZM, Chen Y and Luo ML: Targeting stemness: Implications for precision medicine in breast cancer. Adv Exp Med Biol. 1026:147–169. 2017. View Article : Google Scholar : PubMed/NCBI
10	Fodde R and Brabletz T: Wnt/beta-catenin signaling in cancer stemness and malignant behavior. Curr Opin Cell Biol. 19:150–158. 2007. View Article : Google Scholar : PubMed/NCBI
11	Zhang X, Lou Y, Wang H, Zheng X, Dong Q, Sun J and Han B: Wnt signaling regulates the stemness of lung cancer stem cells and its inhibitors exert anticancer effect on lung cancer SPC-A1 cells. Med Oncol. 32:952015. View Article : Google Scholar : PubMed/NCBI
12	Jin L, Vu T, Yuan G and Datta PK: STRAP promotes stemness of human colorectal cancer via epigenetic regulation of the NOTCH pathway. Cancer Res. 77:5464–5478. 2017. View Article : Google Scholar : PubMed/NCBI
13	Lu T, Li Z, Yang Y, Ji W, Yu Y, Niu X, Zeng Q, Xia W and Lu S: The Hippo/YAP1 pathway interacts with FGFR1 signaling to maintain stemness in lung cancer. Cancer Lett. 423:36–46. 2018. View Article : Google Scholar : PubMed/NCBI
14	Yun Z and Lin Q: Hypoxia and regulation of cancer cell stemness. Adv Exp Med Biol. 772:41–53. 2014. View Article : Google Scholar : PubMed/NCBI
15	Liu X, Li F, Huang Q, Zhang Z, Zhou L, Deng Y, Zhou M, Fleenor DE, Wang H, Kastan MB and Li CY: Self-inflicted DNA double-strand breaks sustain tumorigenicity and stemness of cancer cells. Cell Res. 27:764–783. 2017. View Article : Google Scholar : PubMed/NCBI
16	Carné Trécesson S, Souazé F, Basseville A, Bernard AC, Pécot J, Lopez J, Bessou M, Sarosiek KA, Letai A, Barillé-Nion S, et al: BCL-X_L directly modulates RAS signalling to favour cancer cell stemness. Nat Commun. 8:11232017. View Article : Google Scholar : PubMed/NCBI
17	Sottoriva A, Spiteri I, Piccirillo SG, Touloumis A, Collins VP, Marioni JC, Curtis C, Watts C and Tavaré S: Intratumor heterogeneity in human glioblastoma reflects cancer evolutionary dynamics. Proc Natl Acad Sci USA. 110:4009–4014. 2013. View Article : Google Scholar : PubMed/NCBI
18	Li H, Courtois ET, Sengupta D, Tan Y, Chen KH, Goh JJL, Kong SL, Chua C, Hon LK, Tan WS, et al: Reference component analysis of single-cell transcriptomes elucidates cellular heterogeneity in human colorectal tumors. Nat Genet. 49:708–718. 2017. View Article : Google Scholar : PubMed/NCBI
19	Morgan M, Anders S, Lawrence M, Aboyoun P, Pages H and Gentleman R: ShortRead: A bioconductor package for input, quality assessment and exploration of high-throughput sequence data. Bioinformatics. 25:2607–2608. 2009. View Article : Google Scholar : PubMed/NCBI
20	Bolger AM, Lohse M and Usadel B: Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics. 30:2114–2120. 2014. View Article : Google Scholar : PubMed/NCBI
21	Trapnell C, Cacchiarelli D, Grimsby J, Pokharel P, Li S, Morse M, Lennon NJ, Livak KJ, Mikkelsen TS and Rinn JL: The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells. Nat Biotechnol. 32:381–386. 2014. View Article : Google Scholar : PubMed/NCBI
22	Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, Pimentel H, Salzberg SL, Rinn JL and Pachter L: Differential gene and transcript expression analysis of RNA-seq experiments with tophat and cufflinks. Nat Protoc. 7:562–578. 2012. View Article : Google Scholar : PubMed/NCBI
23	Dalerba P, Cho RW and Clarke MF: Cancer stem cells: Models and concepts. Annu Rev Med. 58:267–284. 2007. View Article : Google Scholar : PubMed/NCBI
24	Wu H, Yu J, Li Y, Hou Q, Zhou R, Zhang N, Jing Z, Jiang M, Li Z, Hua Y, et al: Single-cell RNA sequencing reveals diverse intratumoral heterogeneities and gene signatures of two types of esophageal cancers. Cancer Lett. 438:133–143. 2018. View Article : Google Scholar : PubMed/NCBI
25	Kulsum S, Sudheendra HV, Pandian R, Ravindra DR, Siddappa G, R N, Chevour P, Ramachandran B, Sagar M, Jayaprakash A, et al: Cancer stem cell mediated acquired chemoresistance in head and neck cancer can be abrogated by aldehyde dehydrogenase 1 A1 inhibition. Mol Carcinog. 56:694–711. 2017. View Article : Google Scholar : PubMed/NCBI
26	Liu X, Wang L, Cui W, Yuan X, Lin L, Cao Q, Wang N, Li Y, Guo W, Zhang X, et al: Targeting ALDH1A1 by disulfiram/copper complex inhibits non-small cell lung cancer recurrence driven by ALDH-positive cancer stem cells. Oncotarget. 7:58516–58530. 2016.PubMed/NCBI
27	Meng E, Mitra A, Tripathi K, Finan MA, Scalici J, McClellan S, Madeira da Silva L, Reed E, Shevde LA, Palle K, et al: ALDH1A1 maintains ovarian cancer stem cell-like properties by altered regulation of cell cycle checkpoint and DNA repair network signaling. PLoS One. 9:e1071422014. View Article : Google Scholar : PubMed/NCBI
28	Thomas ML, de Antueno R, Coyle KM, Sultan M, Cruickshank BM, Giacomantonio MA, Giacomantonio CA, Duncan R and Marcato P: Citral reduces breast tumor growth by inhibiting the cancer stem cell marker ALDH1A3. Mol Oncol. 10:1485–1496. 2016. View Article : Google Scholar : PubMed/NCBI
29	Yang L, Ren Y, Yu X, Qian F, Bian BS, Xiao HL, Wang WG, Xu SL, Yang J, Cui W, et al: ALDH1A1 defines invasive cancer stem-like cells and predicts poor prognosis in patients with esophageal squamous cell carcinoma. Mod Pathol. 27:775–783. 2014. View Article : Google Scholar : PubMed/NCBI
30	Chen C, Song G, Xiang J, Zhang H, Zhao S and Zhan Y: AURKA promotes cancer metastasis by regulating epithelial-mesenchymal transition and cancer stem cell properties in hepatocellular carcinoma. Biochem Biophys Res Commun. 486:514–520. 2017. View Article : Google Scholar : PubMed/NCBI
31	Eterno V, Zambelli A, Villani L, Tuscano A, Manera S, Spitaleri A, Pavesi L and Amato A: AurkA controls self-renewal of breast cancer-initiating cells promoting wnt3a stabilization through suppression of miR-128. Sci Rep. 6:284362016. View Article : Google Scholar : PubMed/NCBI
32	Yang N, Wang C, Wang Z, Zona S, Lin SX, Wang X, Yan M, Zheng FM, Li SS, Xu B, et al: FOXM1 recruits nuclear aurora kinase A to participate in a positive feedback loop essential for the self-renewal of breast cancer stem cells. Oncogene. 36:3428–3440. 2017. View Article : Google Scholar : PubMed/NCBI
33	Zheng F, Yue C, Li G, He B, Cheng W, Wang X, Yan M, Long Z, Qiu W, Yuan Z, et al: Nuclear AURKA acquires kinase-independent transactivating function to enhance breast cancer stem cell phenotype. Nat Commun. 7:101802016. View Article : Google Scholar : PubMed/NCBI
34	GursesCila HE, Acar M, Barut FB, Gunduz M, Grenman R and Gunduz E: Investigation of the expression of RIF1 gene on head and neck, pancreatic and brain cancer and cancer stem cells. Clin Invest Med. 39:275002016. View Article : Google Scholar : PubMed/NCBI
35	Li P, Ma X, Adams IR and Yuan P: A tight control of Rif1 by Oct4 and Smad3 is critical for mouse embryonic stem cell stability. Cell Death Dis. 6:e15882015. View Article : Google Scholar : PubMed/NCBI
36	Mei Y, Peng C, Liu YB, Wang J and Zhou HH: Silencing RIF1 decreases cell growth, migration and increases cisplatin sensitivity of human cervical cancer cells. Oncotarget. 8:107044–107051. 2017. View Article : Google Scholar : PubMed/NCBI
37	Li M, Yang J, Zhou W, Ren Y, Wang X, Chen H, Zhang J, Chen J, Sun Y, Cui L, et al: Activation of an AKT/FOXM1/STMN1 pathway drives resistance to tyrosine kinase inhibitors in lung cancer. Br J Cancer. 117:974–983. 2017. View Article : Google Scholar : PubMed/NCBI
38	Obayashi S, Horiguchi J, Higuchi T, Katayama A, Handa T, Altan B, Bai T, Bao P, Bao H, Yokobori T, et al: Stathmin1 expression is associated with aggressive phenotypes and cancer stem cell marker expression in breast cancer patients. Int J Oncol. 51:781–790. 2017. View Article : Google Scholar : PubMed/NCBI
39	Sang Y, Li Y, Song L, Alvarez AA, Zhang W, Lv D, Tang J, Liu F, Chang Z, Hatakeyama S, et al: TRIM59 promotes gliomagenesis by inhibiting TC45 dephosphorylation of STAT3. Cancer Res. 78:1792–1804. 2018. View Article : Google Scholar : PubMed/NCBI
40	Zhou Z, Ji Z, Wang Y, Li J, Cao H, Zhu HH and Gao WQ: TRIM59 is up-regulated in gastric tumors, promoting ubiquitination and degradation of p53. Gastroenterology. 147:1043–1054. 2014. View Article : Google Scholar : PubMed/NCBI
41	Malta TM, Sokolov A, Gentles AJ, Burzykowski T, Poisson L, Weinstein JN, Kamińska B, Huelsken J, Omberg L, Gevaert O, et al: Machine learning identifies stemness features associated with oncogenic dedifferentiation. Cell. 173:338–354.e15. 2018. View Article : Google Scholar : PubMed/NCBI
42	Shibue T and Weinberg RA: EMT, CSCs, and drug resistance: The mechanistic link and clinical implications. Nat Rev Clin Oncol. 14:611–629. 2017. View Article : Google Scholar : PubMed/NCBI
43	Guiu J, Bergen DJ, De Pater E, Islam AB, Ayllon V, Gama-Norton L, Ruiz-Herguido C, González J, López-Bigas N, Menendez P, et al: Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence. J Exp Med. 211:2411–2423. 2014. View Article : Google Scholar : PubMed/NCBI
44	Imai T, Oue N, Sentani K, Sakamoto N, Uraoka N, Egi H, Hinoi T, Ohdan H, Yoshida K, Yasui W, et al: KIF11 is required for spheroid formation by oesophageal and colorectal cancer cells. Anticancer Res. 37:47–55. 2017. View Article : Google Scholar : PubMed/NCBI
45	Jiang M, Zhuang H, Xia R, Gan L, Wu Y, Ma J, Sun Y and Zhuang Z: KIF11 is required for proliferation and self-renewal of docetaxel resistant triple negative breast cancer cells. Oncotarget. 8:92106–92118. 2017. View Article : Google Scholar : PubMed/NCBI
46	Venere M, Horbinski C, Crish JF, Jin X, Vasanji A, Major J, Burrows AC, Chang C, Prokop J, Wu Q, et al: The mitotic kinesin KIF11 is a driver of invasion, proliferation, and self- renewal in glioblastoma. Sci Transl Med. 7:304ra1432015. View Article : Google Scholar : PubMed/NCBI
47	Augustin I, Dewi DL, Hundshammer J, Erdmann G, Kerr G and Boutros M: Autocrine wnt regulates the survival and genomic stability of embryonic stem cells. Sci Signal. 10:eaah68292017. View Article : Google Scholar : PubMed/NCBI
48	Augustin I, Dewi DL, Hundshammer J, Rempel E, Brunk F and Boutros M: Immune cell recruitment in teratomas is impaired by increased wnt secretion. Stem Cell Res. 17:607–615. 2016. View Article : Google Scholar : PubMed/NCBI
49	Ren M and Cowell JK: Constitutive notch pathway activation in murine ZMYM2-FGFR1-induced T-cell lymphomas associated with atypical myeloproliferative disease. Blood. 117:6837–6847. 2011. View Article : Google Scholar : PubMed/NCBI
50	Ren M, Qin H, Wu Q, Savage NM, George TI and Cowell JK: Development of ZMYM2-FGFR1 driven AML in human CD34+ cells in immunocompromised mice. Int J Cancer. 139:836–840. 2016. View Article : Google Scholar : PubMed/NCBI
51	Rubben A and Araujo A: Cancer heterogeneity: Converting a limitation into a source of biologic information. J Transl Med. 15:1902017. View Article : Google Scholar : PubMed/NCBI
52	Mills CC, Kolb EA and Sampson VB: Recent advances of cell-cycle inhibitor therapies for pediatric cancer. Cancer Res. 77:6489–6498. 2017. View Article : Google Scholar : PubMed/NCBI
53	Lord CJ and Ashworth A: PARP inhibitors: Synthetic lethality in the clinic. Science. 355:1152–1158. 2017. View Article : Google Scholar : PubMed/NCBI
54	Weaver AN, Cooper TS, Rodriguez M, Trummell HQ, Bonner JA, Rosenthal EL and Yang ES: DNA double strand break repair defect and sensitivity to poly ADP-ribose polymerase (PARP) inhibition in human papillomavirus 16-positive head and neck squamous cell carcinoma. Oncotarget. 6:26995–277007. 2015. View Article : Google Scholar : PubMed/NCBI
55	Nasuno T, Mimaki S, Okamoto M, Esumi H and Tsuchihara K: Effect of a poly(ADP-ribose) polymerase-1 inhibitor against esophageal squamous cell carcinoma cell lines. Cancer Sci. 105:202–210. 2014. View Article : Google Scholar : PubMed/NCBI
56	Wurster S, Hennes F, Parplys AC, Seelbach JI, Mansour WY, Zielinski A, Petersen C, Clauditz TS, Münscher A, Friedl AA and Borgmann K: PARP1 inhibition radiosensitizes HNSCC cells deficient in homologous recombination by disabling the DNA replication fork elongation response. Oncotarget. 7:9732–9741. 2016. View Article : Google Scholar : PubMed/NCBI
57	Ramalingam SS, Blais N, Mazieres J, Reck M, Jones CM, Juhasz E, Urban L, Orlov S, Barlesi F, Kio E, et al: Randomized, placebo-controlled, phase II study of veliparib in combination with carboplatin and paclitaxel for advanced/metastatic non-small cell lung cancer. Clin Cancer Res. 23:1937–1944. 2017. View Article : Google Scholar : PubMed/NCBI
58	Zhan L, Qin Q, Lu J, Liu J, Zhu H, Yang X, Zhang C, Xu L, Liu Z, Cai J, et al: Novel poly (ADP-ribose) polymerase inhibitor, AZD2281, enhances radiosensitivity of both normoxic and hypoxic esophageal squamous cancer cells. Dis Esophagus. 29:215–223. 2016. View Article : Google Scholar : PubMed/NCBI
59	Yasukawa M, Fujihara H, Fujimori H, Kawaguchi K, Yamada H, Nakayama R, Yamamoto N, Kishi Y, Hamada Y and Masutani M: Synergetic effects of PARP inhibitor AZD2281 and cisplatin in oral squamous cell carcinoma in vitro and in vivo. Int J Mol Sci. 17:2722016. View Article : Google Scholar : PubMed/NCBI