Hypothesis. Hepatocellular carcinoma (HCC) is one of the most malignant human tumors and one of the risk factors is its highly metastatic property. Coptidis Rhizoma aqueous extract (CRAE) is able to suppress the migration and invasion of HCC cells, MHCC97-L, and F-actin reorganization and Rho signaling inhibition is involved. Main methods. CRAE was prepared and analyzed by high-performance liquid chromatography combined with mass spectrometry. The cytotoxicity and antimigration action of CRAE on MHCC97-L cells were evaluated; Immunofluorescence and immunoblotting were used to investigate the proposed mechanism of CRAE action. Key findings . Chemical analysis reveals that the active components in CRAE are berberine and berberine-like alkaloids. CRAE exhibits a significant inhibitory effect on MHCC97-L cell migration as indicated by wound-healing and invasion-chamber assays. No significant alteration of matrix metalloproteinases and urokinase-type plasminogen activator (uPA) expression were observed in MHCC97-L cells exposed to CRAE. Reduction of F-actin polymerization and damage to cytoskeleton network in MHCC97-L cells were observed after CRAE treatment. Furthermore, it was found that CRAE significantly downregulated the Rho/ROCK signaling pathway. Significance. These results indicate that CRAE may act as a Rho/ROCK signaling inhibitor to suppress MHCC97-L cell migration in vitro and suggested that total alkaloids in Coptidis Rhizoma may be a potential agent for suppressing liver cancer invasion.
Gramantieri L., Ferracin M., Fornari F., et al. Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma. Cancer Res. 2007;67: 8-15.
2.
Budhu A., Jia HL, Forgues M., et al. Identification of metastasis-related micrornas in hepatocellular carcinoma. Hepatology. 2008;47:897-907.
3.
Furuse J.Growth factors as therapeutic targets in HCC. Crit Rev Oncol Hematol. 2008;67:8-15.
4.
Tamazaki D., Kurisu S., Takenawa T.Regulation of cancer cell motility through actin reorganization. Cancer Sci. 2005;96:379-386.
5.
Cheng LF, Fung PYScreening for hepatocellular carcinoma: the rationale behind. Hong Kong Pract. 2005;27:469-474.
6.
Chen HW, Lee JY, Huang JYCurcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1. Cancer Res. 2008 ;68:7428-7438.
7.
Hu XW, Meng D., Fang J.Apigenin inhibited migration and invasion of human ovarian cancer A2780 cells through focal adhesion kinase. Carcinogenesis . 2008;29:2369-2376.
8.
Singh RP, Raina K., Sharma G., Agarwal R.Silibinin inhibits established prostate tumor growth, progression, invasion, and metastasis and suppresses tumor angiogenesis and epithelialmesenchymal transition in transgenic adenocarcinoma of the mouse prostate model mice. Clin Cancer Res. 2008;14: 7773-7780.
9.
Kim KS, Rhee HI, Park EK, et al. Anti-inflammatory effects of Radix Gentianae Macrophyllae (Qinjiao), Rhizoma coptidis (huanglian) and Citri Unshiu Pericarpium (Wenzhou migan) in animal models. Chin Med. 2008;3:10.
10.
Kim HY, Shin HS, Park H., et al. In vitro inhibition of coronavirus replications by the traditionally used medicinal herbal extracts, Cimicifuga rhizoma, Meliae cortex, Coptidis rhizoma, and Phellodendron cortex. J Clin Virol. 2008;41: 122-128.
11.
Kwon HA, Kwon YJ, Kwon DY, Lee JHEvaluation of antibacterial effects of a combination of Coptidis Rhizoma, Mume Fructus, and Schizandrae Fructus against Salmonella. Int J Food Microbiol. 2008;127:180-183.
12.
Hwang JM, Wang CJ, Chou FPInhibitory effect of berberine on tert-butyl hydroperoxide-induced oxidative damage in rat liver. Arch Toxicol. 2002;76:664-670.
13.
Luo WQ, Hui SC, Chan TY, Feng Y.Inhibitory effect of water extract from golden thread (huanglian) on leukemia L-1210 cells cultured in vitro. Pharmacologist . 2002;44:A126.
14.
Feng Y., Luo WQ, Zhu SQExplore new clinical application of huanglian and corresponding compound prescriptions from their traditional use [in Chinese]. Zhongguo Zhong Yao Za Zhi. 2008;33:1221-1225.
15.
Tang J., Feng Y., Tsao S., Wang N., Curtain R., Wang Y.Berberine and Coptidis Rhizoma as novel antineoplastic agents: a review of traditional use and biomedical investigations. J Ethnopharmacol . 1999;126:5-17.
16.
Ye X., Feng Y., Tong Y., et al. Hepatoprotective effects of Coptidis Rhizoma aqueous extract on carbon tetrachloride-induced acute liver hepatotoxicity in rats. J Ethnopharmacol. 2009;124:130-136.
17.
Lee TK, Man K., Ho JW, et al. FTY720: a promising agent for treatment of metastatic hepatocellular carcinoma. Clin Cancer Res. 2005;11:8458-8466.
18.
Chen JH, Zhao HQ, Wang XR, Lee SC, Yang HH, Zheng L.Analysis of major alkaloids in Rhizoma coptidis by capillary electrophoresis-electrospray-time of flight mass spectrometry with different background electrolytes. Electrophoresis. 2008;29:2135-2147.
19.
Zhao YG, Xiao AZ, Park HI, et al. Endometase/Matrilysin-2 in human breast ductal carcinoma in situ and its inhibition by tissue inhibitors of metalloproteinases-2 and -4: A putative role in the initiation of breast cancer invasion. Cancer Res. 2004;64:590-598.
20.
Harris CCHepatocellular carcinogenesis: recent advances and speculations. Cancer Cells. 1999;5:146-148.
21.
Kuper H., Ye W., Broomé U., et al. The risk of liver and bile duct cancer in patients with chronic viral hepatitis, alcoholism, or cirrhosis. Hepatology. 2001;34:714-718.
Li Y., Tang ZY, Ye SL, et al. Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol. 2001;7:630-636.
24.
Kang JX, Liu J., Wang J., He C., Li FPThe extract of huanglian, a medicinal herb, induces cell growth arrest and apoptosis by upregulation of interferon-beta and TNFalpha in human breast cancer cells. Carcinogenesis. 2005;26: 1934-1939.
25.
Tsang CM, Lau EP, Di K., et al. Berberine inhibits Rho GTPases and cell migration at low doses but induces G2 arrest and apoptosis at high doses in human cancer cells. Int J Mol Med. 2009;24:131-138.
26.
Olson MF, Sahai E., The actin cytoskeleton cancer cell motility. Clin Exp Metastasis . 2009;26:273-287.
27.
Chen PS, Wang MY, Wu SN, et al. CTGF enhances the motility of breast cancer cells via an integrin-alphavbeta3-ERK1/2-dependent S100A4-upregulated pathway. J Cell Sci. 2007 ;120:2053-2065.
28.
Havaki S., Kouloukoussa M., Amawi K., et al. Altered expression pattern of integrin alphavbeta3 correlates with actin cytoskeleton in primary cultures of human breast cancer. Cancer Cell Int. 2007;7:16.
29.
Smerling C. , Tang K., Hofmann W., Danker K.Role of the alpha(1) integrin cytoplasmic tail in the formation of focal complexes, actin organization, and in the control of cell migration. Exp Cell Res. 2007;313:3153-3165.
30.
Banyard J., Nand-Apte B., Symons M., Zetter BRMotility and invasion are differentially modulated by Rho family GTPases . Oncogene. 2001;19:580-591.
31.
Fritz G., Just I., Kaina B.Rho GTPases are over-expressed in human tumors. Int J Cancer . 1999;81:682-687.
32.
Yoshioka K. , Nakamori S., Itoh K.Overexpression of small GTP-binding protein RhoA promotes invasion of tumor cells. Cancer Res. 2009;59:2004-2010.
33.
Itoh K., Yoshioka K., Akedo H., Uehata M., Ishizaki T., Narumiya S.An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nat Med. 1999;5:221-225.
34.
Takamura M. , Sakamoto M., Genda T., Ichida T., Asakura H., Hirohashi S.Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rho-associated protein kinase inhibitor Y-27632 . Hepatology. 2001;33:577-581.
35.
Wong CC, Wong CM, Ko FC, et al. Deleted in liver cancer 1 (DLC1) negatively regulates Rho/ROCK/MLC pathway in hepatocellular carcinoma. PLoS One. 2008;3:e2779.
36.
Leung TH, Ching YP, Yam JW, et al. Deleted in liver cancer 2 (DLC2) suppresses cell transformation by means of inhibition of RhoA activity. Proc Natl Acad Sci U S A. 2005;102: 15207-15212.
37.
Feng Y., Siu K., Wang N., et al. Bear bile: dilemma of traditional medicinal use and animal protection . J Ethnobiol Ethnomed. 2009;12:2-11.
38.
Iizuka N., Miyamoto K., Okita K., et al. Inhibitory effect of Coptidis Rhizoma and berberine on the proliferation of human esophageal cancer cell lines. Cancer Lett. 2000;148:19-25.
39.
Iizuka N., Oka M., Yamamoto K., et al. Identification of common or distinct genes related to antitumor activities of a medicinal herb and its major component by oligonucleotide microarray. Int J Cancer. 2003;107:666-672.
40.
Li XK, Motwani M., Tong W., Bornmann W., Schwartz GKHuanglian, a Chinese herbal extract, inhibits cell growth by suppressing the expression of cyclin B1 and inhibiting CDC2 kinase activity in human cancer cells. Mol Pharmacol. 2000;58:1287-1293.
41.
Mitani N., Murakami K., Yamaura T., Ikeda T., Saiki I.Inhibitory effect of berberine on the mediastinal lymph node metastasis produced by orthotopic implantation of Lewis lung carcinoma. Cancer Lett. 2001;165:35-42.
42.
Letasiová S., Jantová S., Cipák L., Múcková M.Berberine-antiproliferative activity in vitro and induction of apoptosis/ necrosis of the U937 and B16 cells. Cancer Lett. 2006; 239:254-262.
