HBx promotes hepatoma cell malignant transformation via repressing miR-16 family_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 WU Gang , ZHENG Bo , HUANG Rui , YANG Xun

Department of Hepatobiliary Surgery, Sichuan Provincial People’s Hospital, Chengdu 610072, P.R.China;

Corresponding author：

WU Gang, Email: wugangsamsph@163.com

Keywords：

hepatitis B virus X protein; miRNAs; miR-16 family; c-myc; hepatocellular carcinoma

DOI：

10.7507/1007-9424.201611043

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective Hepatitis B virus X (HBx) protein is involved in the initiation and progression of hepatocellular carcinoma (HCC) by regulating the host protein-coding genes. Herein, we want to explore whether HBx protein can alter the expression of microRNAs (miRNAs) to promote proliferation and transformation in malignant hepatocytesin vitro. Methods MiRNA microarray and quantitative reverse-transcription polymerase chain reactions (qRT-PCRs) were performed to identify miRNAs that were differentially regulated by HBx protein in HCC cells. Protein and mRNA expression analyses, cell cycle and apoptosis analyses, and luciferase reporter assays were performed to delineate the consequences of miR-16 family repression in HepG2 cells. Results HBx protein induced widespread deregulation of miRNAs in HepG2 cells, and the downregulation of the miR-16 family was reproducible in HepG2, SK-HEP-1, and Huh7 cells. CCND1, a target gene of the miR-16 family, was derepressed by HBx protein in HepG2 cells. C-myc mediated the HBx-induced repression of miR-15a/16 in HepG2 cells. Ectopically expressed miR-15a/16 suppressed the proliferation, clonogenicity, and anchorage-independent growth of HBx-expressing HepG2 cells by arresting them in the G₁ phase and inducing apoptosis, whereas reduced expression of miR-16 accelerated the growth and cell-cycle progression of HepG2 cells. Conclusions HBx protein altered thein vitro expression of miRNAs in host malignant hepatocytes, particularly downregulating the miR-16 family. Repression of miR-15a/16 is c-myc mediated and is required for the HBx-induced transformation of HepG2 cellsin vitro. Therefore, miR-16 family may serve as a therapeutic target for hepatitis B virus (HBV)-associated HCC.

Citation： WU Gang, ZHENG Bo, HUANG Rui, YANG Xun. HBx promotes hepatoma cell malignant transformation via repressing miR-16 family. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2017, 24(4): 412-419. doi: 10.7507/1007-9424.201611043 Copy

1.	El-Serag HB. Hepatocellular carcinoma . N Engl J Med, 2011, 365(12): 1118-1127.
2.	Liu S, Koh SS, Lee CG. Hepatitis B virus X protein and hepatocarcinogenesis. Int J Mol Sci, 2016, 17(6). pii: E940.
3.	Lee YS, Dutta A. MicroRNAs in cancer. Annu Rev Pathol, 2009, 4: 199-227.
4.	Calin GA, Croce CM. MicroRNA signatures in human cancers. Nature Rev, 2006, 6(11): 857-866.
5.	Djebali S, Davis CA, Merkel A, et al. Landscape of transcription in human cells. Nature, 2012, 489(7414): 101-108.
6.	Liu Y, Zhao JJ, Wang CM, et al. Altered expression profiles of micrornas in a stable hepatitis B virus-expressing cell line. Chin Med J (Engl), 2009, 122(1): 10-14.
7.	Yue J, Tigyi G. Conservation of miR-15a/16-1 and miR-15b/16-2 clusters. Mamm Genome, 2010, 21(1-2): 88-94.
8.	Linsley PS, Schelter J, Burchard J, et al. Transcripts targeted by the microRNA-16 family cooperatively regulate cell cycle progression. Mol Cell Biol, 2007, 27(6): 2240-2252.
9.	Liu Q, Fu H, Sun F, et al. miR-16 family induces cell cycle arrest by regulating multiple cell cycle genes. Nucleic Acids Res, 2008, 36(16): 5391-5404.
10.	Huang E, Liu R, Chu Y. miRNA-15a/16: as tumor suppressors and more. Future Oncol, 2015, 11(16): 2351-2363.
11.	Blower PE, Chung JH, Verducci JS, et al. MicroRNAs modulate the chemosensitivity of tumor cells. Mol Cancer Ther, 2008, 7(1): 1-9.
12.	Skawran B, Steinemann D, Becker T, et al. Loss of 13q is associated with genes involved in cell cycle and proliferation in dedifferentiated hepatocellular carcinoma. Mod Pathol, 2008, 21(12): 1479-1489.
13.	Wong CM, Lee JM, Lau TC, et al. Clinicopathological significance of loss of heterozygosity on chromosome 13q in hepatocellular carcinoma. Clin Cancer Res, 2002, 8(7): 2266-2272.
14.	Budhu A, Jia HL, Forgues M, et al. Identification of metastasis-related micrornas in hepatocellular carcinoma. Hepatology, 2008, 47(3): 897-907.
15.	Chung GE, Yoon JH, Myung SJ, et al. High expression of microRNA-15b predicts a low risk of tumor recurrence following curative resection of hepatocellular carcinoma. Oncol Rep, 2010, 23(1): 113-119.
16.	Guo CJ, Pan Q, Li DG, et al. miR-15b and miR-16 are implicated in activation of the rat hepatic stellate cell: an essential role for apoptosis. J Hepatol, 2009, 50(4): 766-778.
17.	Guo CJ, Pan Q, Jiang B, et al. Effects of upregulated expression of microRNA-16 on biological properties of culture-activated hepatic stellate cells. Apoptosis, 2009, 14(11): 1331-1340.
18.	Martin-Vilchez S, Sanz-Cameno P, Rodriguez-Munoz Y, et al. The hepatitis B virus X protein induces paracrine activation of human hepatic stellate cells. Hepatology, 2008, 47(6): 1872-1883.
19.	Suzuki HI, Yamagata K, Sugimoto K, et al. Modulation of microRNA processing by p53. Nature, 2009, 460(7254): 529-533.
20.	Bueno MJ, Gomez de Cedron M, Laresgoiti U, et al. Multiple e2f-induced microRNAs prevent replicative stress in response to mitogenic signaling. Mol Cell Biol, 2010, 30(12): 2983-2995.
21.	Chaudhry MA, Sachdeva H, Omaruddin RA. Radiation-induced micro-RNA modulation in glioblastoma cells differing in DNA-repair pathways. DNA Cell Biol, 2010, 29(9): 553-561.
22.	Wagner-Ecker M, Schwager C, Wirkner U, et al. MicroRNA expression after ionizing radiation in human endothelial cells. Radiat Oncol, 2010, 5: 25.
23.	Chang TC, Yu D, Lee YS, et al. Widespread microRNA repression by myc contributes to tumorigenesis. Nat Genet, 2008, 40(1): 43-50.
24.	Kaposi-Novak P, Libbrecht L, Woo HG, et al. Central role of c-myc during malignant conversion in human hepatocarcinogenesis. Cancer Res, 2009, 69(7): 2775-2782.

1. El-Serag HB. Hepatocellular carcinoma . N Engl J Med, 2011, 365(12): 1118-1127.
2. Liu S, Koh SS, Lee CG. Hepatitis B virus X protein and hepatocarcinogenesis. Int J Mol Sci, 2016, 17(6). pii: E940.
3. Lee YS, Dutta A. MicroRNAs in cancer. Annu Rev Pathol, 2009, 4: 199-227.
4. Calin GA, Croce CM. MicroRNA signatures in human cancers. Nature Rev, 2006, 6(11): 857-866.
5. Djebali S, Davis CA, Merkel A, et al. Landscape of transcription in human cells. Nature, 2012, 489(7414): 101-108.
6. Liu Y, Zhao JJ, Wang CM, et al. Altered expression profiles of micrornas in a stable hepatitis B virus-expressing cell line. Chin Med J (Engl), 2009, 122(1): 10-14.
7. Yue J, Tigyi G. Conservation of miR-15a/16-1 and miR-15b/16-2 clusters. Mamm Genome, 2010, 21(1-2): 88-94.
8. Linsley PS, Schelter J, Burchard J, et al. Transcripts targeted by the microRNA-16 family cooperatively regulate cell cycle progression. Mol Cell Biol, 2007, 27(6): 2240-2252.
9. Liu Q, Fu H, Sun F, et al. miR-16 family induces cell cycle arrest by regulating multiple cell cycle genes. Nucleic Acids Res, 2008, 36(16): 5391-5404.
10. Huang E, Liu R, Chu Y. miRNA-15a/16: as tumor suppressors and more. Future Oncol, 2015, 11(16): 2351-2363.
11. Blower PE, Chung JH, Verducci JS, et al. MicroRNAs modulate the chemosensitivity of tumor cells. Mol Cancer Ther, 2008, 7(1): 1-9.
12. Skawran B, Steinemann D, Becker T, et al. Loss of 13q is associated with genes involved in cell cycle and proliferation in dedifferentiated hepatocellular carcinoma. Mod Pathol, 2008, 21(12): 1479-1489.
13. Wong CM, Lee JM, Lau TC, et al. Clinicopathological significance of loss of heterozygosity on chromosome 13q in hepatocellular carcinoma. Clin Cancer Res, 2002, 8(7): 2266-2272.
14. Budhu A, Jia HL, Forgues M, et al. Identification of metastasis-related micrornas in hepatocellular carcinoma. Hepatology, 2008, 47(3): 897-907.
15. Chung GE, Yoon JH, Myung SJ, et al. High expression of microRNA-15b predicts a low risk of tumor recurrence following curative resection of hepatocellular carcinoma. Oncol Rep, 2010, 23(1): 113-119.
16. Guo CJ, Pan Q, Li DG, et al. miR-15b and miR-16 are implicated in activation of the rat hepatic stellate cell: an essential role for apoptosis. J Hepatol, 2009, 50(4): 766-778.
17. Guo CJ, Pan Q, Jiang B, et al. Effects of upregulated expression of microRNA-16 on biological properties of culture-activated hepatic stellate cells. Apoptosis, 2009, 14(11): 1331-1340.
18. Martin-Vilchez S, Sanz-Cameno P, Rodriguez-Munoz Y, et al. The hepatitis B virus X protein induces paracrine activation of human hepatic stellate cells. Hepatology, 2008, 47(6): 1872-1883.
19. Suzuki HI, Yamagata K, Sugimoto K, et al. Modulation of microRNA processing by p53. Nature, 2009, 460(7254): 529-533.
20. Bueno MJ, Gomez de Cedron M, Laresgoiti U, et al. Multiple e2f-induced microRNAs prevent replicative stress in response to mitogenic signaling. Mol Cell Biol, 2010, 30(12): 2983-2995.
21. Chaudhry MA, Sachdeva H, Omaruddin RA. Radiation-induced micro-RNA modulation in glioblastoma cells differing in DNA-repair pathways. DNA Cell Biol, 2010, 29(9): 553-561.
22. Wagner-Ecker M, Schwager C, Wirkner U, et al. MicroRNA expression after ionizing radiation in human endothelial cells. Radiat Oncol, 2010, 5: 25.
23. Chang TC, Yu D, Lee YS, et al. Widespread microRNA repression by myc contributes to tumorigenesis. Nat Genet, 2008, 40(1): 43-50.
24. Kaposi-Novak P, Libbrecht L, Woo HG, et al. Central role of c-myc during malignant conversion in human hepatocarcinogenesis. Cancer Res, 2009, 69(7): 2775-2782.

Previous Article
机器人胃癌手术镜下吻合技术
Next Article
Effect of celecoxib on the expression of NHE1 and intracellular pH in SGC-7901 gastric carcinoma cells

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

HBx promotes hepatoma cell malignant transformation via repressing miR-16 family

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content