Accumulating evidence demonstrated that long non-coding RNAs (lncRNAs) derived from exosomes had the potential to be diagnostic markers for lung cancer. However, the diagnostic value of lncRNAs from epithelial cell adhesion molecule (EpCAM)–positive exosomes remains unclear. In the study, serum EpCAM-positive exosomes were isolated with magnetic beads, and their role in lung cancer was investigated in vitro and in vivo. The copy numbers of lncRNAs RP11-77G23.5 and PHEX-AS1 in EpCAM-specific exosomes were quantified by droplet digital PCR (ddPCR). The diagnostic value of RP11-77G23.5 and PHEX-AS1 was tested in the training cohort and verified in the validation cohort. We found that EpCAM-specific exosomes could promote lung cancer development in vitro and in vivo. RP11-77G23.5 and PHEX-AS1 were significantly elevated in EpCAM-specific exosomes from lung cancer patients and could distinguish malignant from benign lung tumors. The amounts of RP11-77G23.5 were statistically higher in the subtype of lung adenocarcinoma (LUAC) than that of lung squamous cell carcinoma (LUSC), showing its capability to subtype LUAC and LUSC, while PHEX-AS1 exhibited distinct expression signatures between lower and higher tumor stages, and without and with distant metastasis, indicating its association with lung cancer progression. In conclusion, the EpCAM-specific exosomal lncRNAs RP11-77G23.5 and PHEX-AS1 may be promising diagnostic biomarkers for lung cancer.
Serum EpCAM-positive exosomes promote lung cancer development in vitro and in vivo.
Two EpCAM-specific exosomal lncRNAs can be simultaneously detected by RT-ddPCR.
EpCAM-specific exosomal RP11-77G23.5 has the potential to subtype LUAC and LUSC.
EpCAM-specific exosomal PHEX-AS1 is associated with lung cancer progression.
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Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108. https://doi.org/10.3322/caac.21262
Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ (2009) Cancer statistics, 2009. CA Cancer J Clin 59:225–249. https://doi.org/10.3322/caac.20006
Hirsch FR, Franklin WA, Gazdar AF, Bunn PA Jr (2001) Early detection of lung cancer: clinical perspectives of recent advances in biology and radiology. Clin Cancer Res 7:5–22
Duffy MJ, O’Byrne K (2018) Tissue and blood biomarkers in lung cancer: a review. Adv Clin Chem 86:1–21. https://doi.org/10.1016/bs.acc.2018.05.001
Sears CR, Mazzone PJ (2020) Biomarkers in lung cancer. Clin Chest Med 41:115–127. https://doi.org/10.1016/j.ccm.2019.10.004
Paone G, De Angelis G, Munno R, Pallotta G, Bigioni D, Saltini C, Bisetti A, Ameglio F (1995) Discriminant analysis on small cell lung cancer and non-small cell lung cancer by means of NSE and CYFRA-21.1. Eur Respir J 8:1136–1140. https://doi.org/10.1183/09031936.95.08071136
Tanyi JL, Scholler N (2012) Oncology biomarkers for gynecologic malignancies. Front Biosci (Elite Ed) 4:1097–1110. https://doi.org/10.2741/444
Kalluri R, LeBleu VS (2020) The biology, function, and biomedical applications of exosomes. Science 367. https://doi.org/10.1126/science.aau6977
Jalalian SH, Ramezani M, Jalalian SA, Abnous K, Taghdisi SM (2019) Exosomes, new biomarkers in early cancer detection. Anal Biochem 571:1–13. https://doi.org/10.1016/j.ab.2019.02.013
Meng XD, Pan JC, Sun SF, Gong ZH (2018) Circulating exosomes and their cargos in blood as novel biomarkers for cancer. Translational Cancer Research 7:S226–S242. https://doi.org/10.21037/tcr.2017.09.17
Yang YF, Sun XY, Shen XT, Ye GX, Lou CT, Li N, Gong ZH, Meng XD (2020) The role of exosomal MiRNAs in the occurrence and development of lung cancer. Progress in Biochemistry and Biophysics 47:188–198. https://doi.org/10.16476/j.pibb.2019.0273
Mashouri L, Yousefi H, Aref AR, Ahadi AM, Molaei F, Alahari SK (2019) Exosomes: composition, biogenesis, and mechanisms in cancer metastasis and drug resistance. Mol Cancer 18:75. https://doi.org/10.1186/s12943-019-0991-5
Li K, Chen Y, Li A, Tan C, Liu X (2019) Exosomes play roles in sequential processes of tumor metastasis. Int J Cancer 144:1486–1495. https://doi.org/10.1002/ijc.31774
Gires O, Pan M, Schinke H, Canis M, Baeuerle PA (2020) Expression and function of epithelial cell adhesion molecule EpCAM: where are we after 40 years? Cancer Metastasis Rev 39:969–987. https://doi.org/10.1007/s10555-020-09898-3
Yahyazadeh Mashhadi SM, Kazemimanesh M, Arashkia A, Azadmanesh K, Meshkat Z, Golichenari B, Sahebkar A (2019) Shedding light on the EpCAM: an overview. J Cell Physiol 234:12569–12580. https://doi.org/10.1002/jcp.28132
Meng X, Müller V, Milde-Langosch K, Trillsch F, Pantel K, Schwarzenbach H (2016) Diagnostic and prognostic relevance of circulating exosomal miR-373, miR-200a, miR-200b and miR-200c in patients with epithelial ovarian cancer. Oncotarget 7:16923–16935. https://doi.org/10.18632/oncotarget.7850
Li J, Tian H, Pan J, Jiang N, Yang J, Zhou C, Xu D, Meng X, Gong Z (2017) Pecanex functions as a competitive endogenous RNA of S-phase kinase associated protein 2 in lung cancer. Cancer Lett 406:36–46. https://doi.org/10.1016/j.canlet.2017.07.030
Bloom AC, Bender LH, Tiwary S, Pasquet L, Clark K, Jiang T, Xia Z, Morales-Kastresana A, Jones JC, Walters I et al (2019) Intratumorally delivered formulation, INT230-6, containing potent anticancer agents induces protective T cell immunity and memory. Oncoimmunology 8:e1625687. https://doi.org/10.1080/2162402x.2019.1625687
Jin X, Chen Y, Chen H, Fei S, Chen D, Cai X, Liu L, Lin B, Su H, Zhao L et al (2017) Evaluation of tumor-derived exosomal miRNA as potential diagnostic biomarkers for early-stage non-small cell lung cancer using next-generation sequencing. Clin Cancer Res 23:5311–5319. https://doi.org/10.1158/1078-0432.ccr-17-0577
Hannafon BN, Trigoso YD, Calloway CL, Zhao YD, Lum DH, Welm AL, Zhao ZJ, Blick KE, Dooley WC, Ding WQ (2016) Plasma exosome microRNAs are indicative of breast cancer. Breast Cancer Res 18:90. https://doi.org/10.1186/s13058-016-0753-x
Lee YR, Kim G, Tak WY, Jang SY, Kweon YO, Park JG, Lee HW, Han YS, Chun JM, Park SY et al (2019) Circulating exosomal non-coding RNAs as prognostic biomarkers in human hepatocellular carcinoma. Int J Cancer 144:1444–1452. https://doi.org/10.1002/ijc.31931
Jiang N, Pan J, Fang S, Zhou C, Han Y, Chen J, Meng X, Jin X, Gong Z (2019) Liquid biopsy: circulating exosomal long non-coding RNAs in cancer. Clin Chim Acta 495:331–337. https://doi.org/10.1016/j.cca.2019.04.082
Zhu L, Li J, Gong Y, Wu Q, Tan S, Sun D, Xu X, Zuo Y, Zhao Y, Wei YQ et al (2019) Exosomal tRNA-derived small RNA as a promising biomarker for cancer diagnosis. Mol Cancer 18:74. https://doi.org/10.1186/s12943-019-1000-8
Zhang X, Shi H, Yuan X, Jiang P, Qian H, Xu W (2018) Tumor-derived exosomes induce N2 polarization of neutrophils to promote gastric cancer cell migration. Mol Cancer 17:146. https://doi.org/10.1186/s12943-018-0898-6
Razzo BM, Ludwig N, Hong CS, Sharma P, Fabian KP, Fecek RJ, Storkus WJ, Whiteside TL (2020) Tumor-derived exosomes promote carcinogenesis of murine oral squamous cell carcinoma. Carcinogenesis 41:625–633. https://doi.org/10.1093/carcin/bgz124
van der Gun BT, Melchers LJ, Ruiters MH, de Leij LF, McLaughlin PM, Rots MG (2010) EpCAM in carcinogenesis: the good, the bad or the ugly. Carcinogenesis 31:1913–1921. https://doi.org/10.1093/carcin/bgq187
Baeuerle PA, Gires O (2007) EpCAM (CD326) finding its role in cancer. Br J Cancer 96:417–423. https://doi.org/10.1038/sj.bjc.6603494
Zhou Y, Wang B, Wu J, Zhang C, Zhou Y, Yang X, Zhou J, Guo W, Fan J (2016) Association of preoperative EpCAM circulating tumor cells and peripheral Treg cell levels with early recurrence of hepatocellular carcinoma following radical hepatic resection. BMC Cancer 16:506. https://doi.org/10.1186/s12885-016-2526-4
Coumans FA, Ligthart ST, Uhr JW, Terstappen LW (2012) Challenges in the enumeration and phenotyping of CTC. Clin Cancer Res 18:5711–5718. https://doi.org/10.1158/1078-0432.ccr-12-1585
Chen G, Huang AC, Zhang W, Zhang G, Wu M, Xu W, Yu Z, Yang J, Wang B, Sun H et al (2018) Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response. Nature 560:382–386. https://doi.org/10.1038/s41586-018-0392-8
Theodoraki MN, Yerneni SS, Hoffmann TK, Gooding WE, Whiteside TL (2018) Clinical significance of PD-L1(+) exosomes in plasma of head and neck cancer patients. Clin Cancer Res 24:896–905. https://doi.org/10.1158/1078-0432.ccr-17-2664
Xie F, Xu M, Lu J, Mao L, Wang S (2019) The role of exosomal PD-L1 in tumor progression and immunotherapy. Mol Cancer 18:146. https://doi.org/10.1186/s12943-019-1074-3
Schwarzenbach H, da Silva AM, Calin G, Pantel K (2015) Data normalization strategies for MicroRNA quantification. Clin Chem 61:1333–1342. https://doi.org/10.1373/clinchem.2015.239459
We thank Prof. Junming Guo for providing us with the ddPCR instruments.
The study was partially supported by a grant from the National Natural Science Foundation of China (81902146), the Natural Science Foundation of Zhejiang Province (LQ18H200001), the Program of “Xinmiao” (Potential) Talents in Zhejiang Province (2020R405041, 2021R405044), the Research Project in Ningbo University (JYXMXZD2021031, XYL20028), the Research Project of Education Science in Ningbo City (2020YGH007) and the K.C.Wong Magna Fund in Ningbo University.
Ethics approval and consent to participate
All animal experiments were performed in accordance with currently prescribed guidelines and followed a protocol approved by the Medical School of Ningbo University. This study was conducted in accordance with the Declaration of Helsinki. Written informed consent was obtained from all patients, and the study was approved by the ethics committee (the Clinical Research Ethics Committee of Medical School of Ningbo University).
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Shen, X., Yang, Y., Chen, Y. et al. Evaluation of EpCAM-specific exosomal lncRNAs as potential diagnostic biomarkers for lung cancer using droplet digital PCR. J Mol Med (2021). https://doi.org/10.1007/s00109-021-02145-4
- EpCAM-specific exosomes
- Lung cancer
- Lung adenocarcinoma
- Squamous cell lung cancer
- Digital polymerase chain reaction