Information to authors
Identification and differential expression of microRNA in response to elevated phospholipase Cγ expression in liver RH 35 carcinoma cells
Our study has primarily shown the positive effect of PLCγ2 on liver tumor cell proliferation, but the molecular basis for its function remains elusive. miRNAs have been widely accepted as important modulators of various cellular activities. This study attempts to characterize the global influence of PLCγ2 on miRNA expressions in liver cancer RH35 cells. Firstly, the recombinant adenovirus AdPLCγ2 was infected into the cells. Highthroughput sequencing technology was applied to measure miRNA expressions in PLCγ2overexpressing cells. Moreover, the target genes and signaling pathways modulated by PLCγ2specifc miRNAs were identified using target prediction program, GO annotation and KEGG analysis. As a result, totally 246 known and 1075 novel candidate miRNAs were identified, among which 34 known and 191 novel miRNAs exhibited ≥ 2fold changes in the AdPLCγ2infected cells. Correspondingly, 6985 target genes of above 225 differentlyexpressed miRNAs were predicted, mainly involved in Hippo signaling, Wnt signaling etc., and responsible for tumor development, cell proliferation, apoptosis, migration, lipid metabolism and so on. In aggregate, PLCγ2 induces the significant alterations in miRNA expression, thus providing mechanistic insights into tumorgenesis mediated by PLCγ2, and maybe offers some clues on identifying potential candidates for controlling liver cancer cell growth.
Key words: Phospholipase Cγ2, liver carcinoma, highthroughput sequencing, microRNA expression, target prediction
E-mail: cxgtzr2012 163.com
1. Regad, T., Targeting RTK signaling pathways in cancer, Cancers (Basel), 2015, vol. 7, pp. 1758–1784.
2. Browaeys-Poly, E., Perdereau, D., Lescuyer, A., Burnol, A.F., and Cailliau, K., Akt interaction with PLC(gamma) regulates the G(2)/M transition triggered by FGF receptors from MDA-MB-231 breast cancer cells, Anticancer Res., 2009, vol. 29, no. 12, pp. 4965–4969.
3. Zhang, P., Zhao, Y, Zhu, X., Sedwick, D., Zhang, X., and Wang, Z., Cross-talk between phospho-STAT3 and PLCγ1 plays a critical role in colorectal tumorigenesis, Mol. Cancer Res., 2011, vol. 9, no. 10, pp. 1418–1428.
4. Khoshyomn, S., Penar, P.L., Rossi, J., Wells, A., Abramson, D.L., and Bhushan, A., Inhibition of phospholipase C-gammal activation blocks glioma cell motility and invasion of fetal rat brain aggregates, Neurosurgery, 1999, vol. 44, no. 3, pp. 568–578.
5. Koss, H., Bunney, T.D., Behjati, S., and Katan, M, Dysfunction of phospholipase Cγ in immune disorders and cancer, Trends Biochem Set., 2014, vol. 39, no. 12, pp. 603–611.
6. Tensen, CP, PLCG1 gene mutations in cutaneous T-cell lymphomas revisited, J. Invest. Dermatol., 2015, vol. 135, no. 9, pp. 2153–2154.
7. Feng, L., Reynisdóttir, I., and Reynisson, J., The effect of PLC-γ2 inhibitors on the growth of human tumour cells, Eur J. Med. Chem., 2012, vol. 54, pp. 463–469.
8. Huynh, M.Q., Goẞmann, J., Gattenlöehner, S., Klapper, W., Wacker, H.H., Ramaswamy, A., Bittner, A., Kaiser, U., and Neubauer, A., Expression and pro-survival function of phospholipase Cγ2 in diffuse large B-cell lymphoma, Leuk. Lymphoma, 2015, vol. 56, no. 4, pp. 1088–1095.
9. Liu, T.M., Woyach, J.A., Zhong, Y., Lozanski, A., Lozanski, G, Dong, S., Strattan, E., Lehman, A., Zhang, X., Jones, J.A., Flynn, J., Andritsos, L.A., Maddocks, K., Jaglowski, S.M., Blum, K.A., Byrd, J.C., Dubovsky, J.A., and Johnson, A.J., Hypermorphic mutation of phospholipase C, γ2 acquired in ibrutinib-resistant CLL confers BTK independency upon B-cell receptor activation, Blood, 2015, vol. 126, no. 1, pp. 61–68.
10. Ghouri, Y.A., Mian, I., and Rowe, J.H., Review of hepatocellular carcinoma: Epidemiology, etiology, and carcinogenesis, J. Carcinog., 2017, vol. 16, p. 1.
11. Gramantieri, L., Fornari, F, Callegari, E., Sabbioni, S., Lanza, G, Croce, C.M., Bolondi, L., and Negrini, M., MicroRNA involvement in hepatocellular carcinoma, J. Cell Mol. Med., 2008, vol. 12, no. 6A, pp. 2189–204.
12. Aravalli, R.N., Cressman, E.N., and Steer, C.J., Cellular and molecular mechanisms of hepatocellular carcinoma: an update, Arch. Toxicol., 2013, vol. 87, no. 2, pp. 227–247.
13. Lee, J.S., Chu, I.S., Heo, J., Calvisi, D.F, Sun, Z., Roskams, T., Durnez, A., Demetris, A.J., and Thorgeirsson, S.S., Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling, Hepatology, 2004, vol. 40, no. 3, pp. 667–676.
14. Ji, J., Shi, J., Budhu, A., Yu, Z., Forgues, M., Roessler, S., Ambs, S., Chen, Y., Meltzer, P.S., Cãoce, C.M., Qin, L.X., Man, K., Lo, CM., Lee, J., Ng, I.O., Fan, J., Tang, Z.Y., Sun, H.C., and Wang, X.W., MicroRNA expression, survival, and response to interferon in liver cancer, N. Engl. J. Med., 2009, vol. 361, no. 15, pp. 1437–1447.
15. Ranganathan, K., Sivasankar, V., microRNAs-Biology and clinical applications, J. Oral. Maxillofac. Pathol., 2014, vol. 18, no. 2, pp. 229–234.
16. Esquela-Kerscher, A., Slack, F.J., Oncomirs-microRNAs with a role in cancer, Nat. Rev. Cancer, 2006, vol. 6, no. 4, pp. 259–269.
17. Callegari, E., Gramantieri, L., Domenicali, M., Dabundo, L., Sabbioni, S., and Negrini, M., MicroRNAs in liver cancer: a model for investigating pathogenesis and novel therapeutic approaches, Cell Death. Differ., 2015, vol. 22, no. 1, pp. 46–57.
18. Shah, M., Calin, G.A., MicroRNAs as therapeutic targets in human cancers, Wiley Interdiscip. Rev. RNA, 2014, vol. 5, no. 4, pp. 537–548.
19. Gautam, A., Kumar, R., Dimitrov, G, Hoke, A., Hammamieh, R., and Jett, M., Identification of extracellular miRNA in archived serum samples by next-generation sequencing from RNA extracted using multiple methods, Mol. Biol. Rep., 2016, vol. 43, no. 10, pp. 1165–1178.
20. Gyvyte, U., Juzenas, S., Salteniene, V., Kupcinskas, J., Poskiene, L., Kucinskas, L., Jarmalaite, S., Stuopelyte, K., Steponaitiene, R., Hemmrich-Stanisak, G., Hübenthal, M., Link, A., Franke, S., Franke, A., Pangonyte, D., Lesauskaite, V., Kupcinskas, L., and Skieceviciene, J., MiRNA profiling of gastrointestinal stromal tumors by next-generation sequencing, Oncotarget, 2017, vol. 8, no. 23, pp. 37225–37238.
21. Chen, X., Lv, Q., Liu, Y., and Deng, W., Construction of recombinant adenovirus Ad-rat PLCγ2 and its effects on apoptosis of rat liver cell BRL-3A in vitro, Cell Mol. Biol. (Noisy-le-grand), 2016, vol. 62, no. 11, pp. 45–50.
22. Chen, X., Lv, Q., Ma, J., and Liu,Y., PLCγ2 promotes apoptosis while inhibits proliferation in rat hepatocytes through PKCD/JNK ÌÀÐÊ and PKCD/p38 ÌÀÐÊ signaling, Cell Prolif., 2018, vol. 51, no. 3, p. e²2437.
23. Martin, M., Cutadapt removes adapter sequences from high-throughput sequencing reads, EMBnet J., 2011, vol. 17, pp. 10–12.
24. Friedländer, M.R., Chen, W., Adamidi, C, Maaskola, J., Einspanier, R., Knespel, S., and Rajewsky, N., Discovering microRNAs from deep sequencing data using miRDeep, Nat. Biotechnol., 2008, vol. 26, no. 4, pp. 407–415.
25. Anders, S., Huber, W., Differential expression analysis for sequence count data, Genome Biol., 2010, vol. 11, no. 10, p. R106.
26. John, B., Enright, A.J., Aravin, A., Tuschl, T., Sander, C., and Marks, D.S., Human microRNA targets, PLoS Biol., 2005, vol. 3, no. 7, p. e264.
27. Ye, J., Zhang, Y., Cui, H., Liu, J., Wu, Y., Cheng, Y., Xu, H., Huang, X., Li, S., Zhou, A., Zhang, X., Bolund, L., Chen, Q., Wang, J., Yang, H., Fang, L., and Shi, C., WEGO 2.0: a web tool for analyzing and plotting GO annotations, 2018 update, Nucleic Acids Res., 2018, vol. 46, no. W1, pp. W71–W75.
28. Zhou, K., Liu, M., and Cao, Y., New Insight into microRNA functions in cancer: oncogene-microRNA–tumor suppressor gene network, Front. Mol. Biosci., 2017, vol. 4, p. 46.
29. Tang, W., Wan, S., Yang, Z., Teschendorff, A.E., and Zou, Q., Tumor origin detection with tissue-specific miRNA and DNA methylation markers, Bioinformatics, 2018, vol. 34, no. 3, pp. 398–406.
30. Li, D.B., Liu, J.L., Wang, W., Luo, X.M., Zhou, X., Li, J.P., Cao, X.L., Long, X.H., Chen, J.G., and Qin, C., Plasma exosomal miRNA-122-5p and miR-300-3p as potential markers for transient ischaemic attack in rats, Front. Aging. Neurosci., 2018, vol. 10, p. 24.
31. Nour, M., Scalzo, F., and Liebeskind, D.S., Ischemia–reperfusion injury in stroke, Interv. Neurol., 2013, vol. 1, no. 3–4, pp. 85–199.
32. Takuma, A., Abe, A., Saito, Y., Nito, C., Ueda, M., Ishimaru, Y., Harada, H., Abe, K., Kimura, K., and Asakura, T., Gene expression analysis of the effect of ischemic infarction in whole blood, Int. J. Mol. Sci., 2017, vol. 18, no. 11, p.E2335.
33. Chen, F., Wang, R.J., Li, G.Z., Zhang, Y., Yu, S., Liu, Y.F., Chen, X.Y., and Hou, S.K., miRNA array analysis of plasma miRNA alterations in rats exposed to a high altitude hypoxic environment, Mol. Med. Rep., 2018, vol. 18, no. 6, pp. 5502–5510.
34. Tong, Y.J., miRNA expression analysis of effect of aerobic exercise on apoptosis of spermatogenic cells in high-fat diet rats, Yangzhou Univ., 2017.
35. Oliveto, S., Mancino, M., Manfrini, N., and Biffo, S., Role of microRNAs in translation regulation and cancer, World J. Biol. Chem., 2017, vol. 8, no. 1, pp. 45–56.
36. Chen, C., Wells, A.D., Comparative analysis of E2F family member oncogenic activity, PLoS One, 2007, vol. 2, p. e912.
37. Opavsky, R., Tsai, S.Y., Guimond, M., Arora, A., Opavska, J., Becknell, B., Kaufmann, M., Walton, N.A., Stephens, J.A., Fernandez, S.A., Muthusamy, N., Felsher, D.W., Porcu, P., Caligiuri, M.A., and Leone, G., Specific tumor suppressor function for E2F2 in Myc-induced T cell lymphomagenesis, Proc. Natl. Acad. Sci. U. S. A., 2007, vol. 104, no. 39, pp. 15400–15405.
38. Warren, J.S.A., Xiao, Y., and Lamar, J.M., YAP/TAZ activation as a target for treating metastatic cancer, Cancers (Basel), 2018, vol. 10, no. 4, p. E115.
39. Khalaf, A.M., Fuentes, D., Morshid, A.I., Burke, M.R., Kaseb, A.O., Hassan, M., Hazle, J.D., and Elsayes, K.M., Role of Wnt/β-catenin signaling in hepatocellular carcinoma, pathogenesis, and clinical significance, J. Hepatocell. Carcinoma, 2018, vol. 5, pp. 61–73.
40. Scharenberg, A.M., Humphries, L.A., and Rawlings, D.J., Calcium signalling and cell-fate choice in B cells, Nat. Rev. Immunol., 2007, vol. 7, no. 10, pp. 778–789.
|Coded & Designed by Volodymyr Duplij||Modified 24.09.21|