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Knockdown of MACC1 expression suppressed hepatocellular carcinoma cell migration and invasion and inhibited expression of MMP2 and MMP9

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Abstract

Expression of MACC1 (metastasis-associated in colon cancer-1) protein is associated with metastasis of various human cancers. This study analyzed MACC1 protein expression in hepatocellular carcinoma (HCC) tissue specimens and then investigated the effects of MACC1 knockdown on HCC cell migration and invasion, and gene expression levels. Sixty pairs of HCC and adjacent normal liver tissues from HCC patients were analyzed for MACC1 expression immunohistochemically. The HCC cell lines Hep3B, Huh7, MHCC97H, SMMC-7721, Bel-7402, and HepG2 and the normal liver cell line LO2 were used to assess expressions of MACC1 mRNA and MACC1 protein using qRT-PCR and western blot, respectively. MACC1 short hairpin RNA (shRNA) was used to knockdown MACC1 protein expression in Huh7 cells. Changes in the tumor phenotype of these cells were analyzed with wound healing assay and invasion assays, and differences in gene expression were evaluated via western blot. Immunofluorescence was used to locate MACC1 protein in the above cell lines. MACC1 was highly expressed in HCC tissues and the nuclear expression of MACC1 protein was associated with poor tumor differentiation and intrahepatic metastasis or portal invasion. Moreover, MACC1 mRNA and MACC1 protein was also expressed in HCC cell lines. Immunostaining showed that MACC1 protein was localized in both nuclei and cytoplasm of HCC cell lines and the nuclear localization of MACC1 protein was associated with increased aggressiveness of HCC in cell lines. Knockdown of MACC1 expression using MACC1-shRNA reduced Huh7 cell migration and invasion abilities, which was associated with downregulation of MMP2, MMP9, and c-Met proteins in Huh7 cells. Localization of MACC1 protein to the nucleus may predict HCC progression. Knockdown of MACC1 expression using MACC1 shRNA warrants further evaluation as a novel therapeutic strategy for control of HCC.

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Abbreviations

c-Met:

Hepatocyte growth factor receptor

HCC:

Hepatocellular carcinoma

MACC1:

Metastasis-associated colon cancer 1

MMP2/9:

Matrix metalloproteinase 2/9

References

  1. Jemal A, Bray F, Center MM et al (2011) Global cancer statistics. CA Cancer J Clin 61(2):69–90

    Article  PubMed  Google Scholar 

  2. Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257(5072):967–971

    Article  PubMed  CAS  Google Scholar 

  3. Stein U, Walther W, Arlt F et al (2009) MACC1, a newly identified key regulator of HGF-MET signaling, predicts colon cancer metastasis. Nat Med 15(1):59–67

    Article  PubMed  CAS  Google Scholar 

  4. Chundong G, Uramoto H, Onitsuka T et al (2011) Molecular diagnosis of MACC1 status in lung adenocarcinoma by immunohistochemical analysis. Anticancer Res 31(4):1141–1145

    PubMed  Google Scholar 

  5. Stein U, Dahlmann M, Walther W (2010) MACC1—more than metastasis? Facts and predictions about a novel gene. J Mol Med (Berl) 88(1):11–18

    Article  CAS  Google Scholar 

  6. Arlt F, Stein U (2009) Colon cancer metastasis: MACC1 and Met as metastatic pacemakers. Int J Biochem Cell Biol 41(12):2356–2359

    Article  PubMed  CAS  Google Scholar 

  7. Shirahata A, Fan W, Sakuraba K et al (2011) MACC 1 as a marker for vascular invasive hepatocellular carcinoma. Anticancer Res 31(3):777–780

    PubMed  Google Scholar 

  8. Qiu J, Huang P, Liu Q et al (2011) Identification of MACC1 as a novel prognostic marker in hepatocellular carcinoma. J Transl Med 9:166

    Article  PubMed  CAS  Google Scholar 

  9. Kokoszynska K, Krynski J, Rychlewski L et al (2009) Unexpected domain composition of MACC1 links MET signaling and apoptosis. Acta Biochim Pol 56(2):317–323

    PubMed  CAS  Google Scholar 

  10. Weidner KM, Arakaki N, Hartmann G et al (1991) Evidence for the identity of human scatter factor and human hepatocyte growth factor. Proc Natl Acad Sci USA 88(16):7001–7005

    Article  PubMed  CAS  Google Scholar 

  11. Bardelli A, Comoglio PM (1997) Scatter factor receptors are key players in a unique multistep program leading to invasive growth. Ciba Found Symp 212:133–144 (discussion 144–137)

    Google Scholar 

  12. Baldus SE, Kort EJ, Schirmacher P et al (2007) Quantification of MET and hepatocyte growth factor/scatter factor expression in colorectal adenomas, carcinomas and non-neoplastic epithelia by quantitative laser scanning microscopy. Int J Oncol 31(1):199–204

    PubMed  CAS  Google Scholar 

  13. Birchmeier C, Birchmeier W, Gherardi E et al (2003) Met, metastasis, motility and more. Nat Rev Mol Cell Biol 4(12):915–925

    Article  PubMed  CAS  Google Scholar 

  14. Stein U, Smith J, Walther W et al (2009) MACC1 controls Met: what a difference an Sp1 site makes. Cell Cycle 8(15):2467–2469

    Article  PubMed  CAS  Google Scholar 

  15. Sattler M, Salgia R (2007) c-Met and hepatocyte growth factor: potential as novel targets in cancer therapy. Curr Oncol Rep 9(2):102–108

    Article  PubMed  CAS  Google Scholar 

  16. Bottaro DP, Rubin JS, Faletto DL et al (1991) Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. Science 251(4995):802–804

    Article  PubMed  CAS  Google Scholar 

  17. Chmielowiec J, Borowiak M, Morkel M et al (2007) c-Met is essential for wound healing in the skin. J Cell Biol 177(1):151–162

    Article  PubMed  CAS  Google Scholar 

  18. Kaposi-Novak P, Lee JS, Gomez-Quiroz L et al (2006) Met-regulated expression signature defines a subset of human hepatocellular carcinomas with poor prognosis and aggressive phenotype. J Clin Invest 116(6):1582–1595

    Article  PubMed  CAS  Google Scholar 

  19. Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2(3):161–174

    Article  PubMed  CAS  Google Scholar 

  20. Bjorklund M, Koivunen E (2005) Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta 1755(1):37–69

    PubMed  Google Scholar 

  21. Canel M, Secades P, Garzon-Arango M et al (2008) Involvement of focal adhesion kinase in cellular invasion of head and neck squamous cell carcinomas via regulation of MMP-2 expression. Br J Cancer 98(7):1274–1284

    Article  PubMed  CAS  Google Scholar 

  22. Lamar JM, Pumiglia KM, DiPersio CM (2008) An immortalization-dependent switch in integrin function up-regulates MMP-9 to enhance tumor cell invasion. Cancer Res 68(18):7371–7379

    Article  PubMed  CAS  Google Scholar 

  23. Zheng H, Takahashi H, Murai Y et al (2006) Expressions of MMP-2, MMP-9 and VEGF are closely linked to growth, invasion, metastasis and angiogenesis of gastric carcinoma. Anticancer Res 26(5A):3579–3583

    PubMed  CAS  Google Scholar 

  24. Qu JH, Chang XJ, Lu YY et al (2012) Overexpression of metastasis-associated in colon cancer 1 predicts a poor outcome of hepatitis B virus-related hepatocellular carcinoma. World J Gastroenterol 18(23):2995–3003

    Article  PubMed  CAS  Google Scholar 

  25. Sainz B Jr, Barretto N, Uprichard SL (2009) Hepatitis C virus infection in phenotypically distinct Huh7 cell lines. PLoS ONE 4(8):e6561

    Article  PubMed  Google Scholar 

  26. Shirahata A, Shinmura K, Kitamura Y et al (2010) MACC1 as a marker for advanced colorectal carcinoma. Anticancer Res 30(7):2689–2692

    PubMed  CAS  Google Scholar 

  27. Shirahata A, Sakata M, Kitamura Y et al (2010) MACC 1 as a marker for peritoneal-disseminated gastric carcinoma. Anticancer Res 30(9):3441–3444

    PubMed  Google Scholar 

  28. Shimokawa H, Uramoto H, Onitsuka T et al (2011) Overexpression of MACC1 mRNA in lung adenocarcinoma is associated with postoperative recurrence. J Thorac Cardiovasc Surg 141(4):895–898

    PubMed  CAS  Google Scholar 

  29. Deryugina EI, Quigley JP (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25(1):9–34

    Article  PubMed  CAS  Google Scholar 

  30. Fingleton B (2006) Matrix metalloproteinases: roles in cancer and metastasis. Front Biosci 11:479–491

    Article  PubMed  CAS  Google Scholar 

  31. Kessenbrock K, Plaks V, Werb Z (2010) Matrix metalloproteinases: regulators of the tumor microenvironment. Cell 141(1):52–67

    Article  PubMed  CAS  Google Scholar 

  32. Bu W, Huang X, Tang Z (1997) The role of MMP-2 in the invasion and metastasis of hepatocellular carcinoma (HCC). Zhonghua Yi Xue Za Zhi 77(9):661–664

    PubMed  CAS  Google Scholar 

  33. Yang P, Yuan W, He J et al (2009) Overexpression of EphA2, MMP-9, and MVD-CD34 in hepatocellular carcinoma: implications for tumor progression and prognosis. Hepatol Res 39(12):1169–1177

    Article  PubMed  Google Scholar 

  34. Edmondson HA, Steiner PE (1954) Primary carcinoma of the liver: a study of 100 cases among 48,900 necropsies. Cancer 7(3):462–503

    Article  PubMed  CAS  Google Scholar 

  35. Zhou L, Rui JA, Ye DX et al (2008) Edmondson-Steiner grading increases the predictive efficiency of TNM staging for long-term survival of patients with hepatocellular carcinoma after curative resection. World J Surg 32(8):1748–1756

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Medjaden Bioscience Limited, Hong Kong, China, for assisting in the preparation of this manuscript. This study was supported in part by grants from the National Natural Scientific Foundation of China (No. 81071897 to YY).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, 277 West Yanta Road, Xi’an, China

    Jie Gao, Feihu Ding, Qingguang Liu & Yingmin Yao

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  1. Jie Gao
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  2. Feihu Ding
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  3. Qingguang Liu
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  4. Yingmin Yao
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Correspondence to Qingguang Liu or Yingmin Yao.

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11010_2012_1545_MOESM1_ESM.tif

Supplement Figure 1: Knockdown of MACC1 mRNA expression by MACC1-1 and MACC1-2 shRNA transfection. The HCC cells were grown and transiently transfected with MACC1-1 and MACC1-2 siRNA for 24 h or 48 h and total cellular RNA was isolated and subjected to RT-PCR analysis (TIFF 88 kb)

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Gao, J., Ding, F., Liu, Q. et al. Knockdown of MACC1 expression suppressed hepatocellular carcinoma cell migration and invasion and inhibited expression of MMP2 and MMP9. Mol Cell Biochem 376, 21–32 (2013). https://doi.org/10.1007/s11010-012-1545-y

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  • DOI: https://doi.org/10.1007/s11010-012-1545-y

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