인체 간암세포에서 β-lapachone 처리에 의한 Tight Junction 관련 유전자의 변화 = (β-lapachone Regulates Tight Junction Proteins, Claudin-3 and -4, in Human Hepatocarcinoma Cells.

국문 초록 (Abstract)

${\beta}-lapachone$은 남미지역에서 자생하는 Tabebuia avellanedae라는 나무의 수피에서 동정된 quinone계 물질로서 다양한 인체암세포에서 항암효과가 있는 것으로 알려져있다. 본 연구에서는 ${\beta}-lapachone$의 암 전이 억제에 대한 연구의 일환으로 HepG2 및 Hep3B 인체 간암 세포의 전이관련 유전자의 발현에 미치는 ${\beta}-lapachone$의 영향을 조사하였다. MTT assay 및 세포형태변화 관찰 결과에서 ${\beta}-lapachone$ 처리에 따라 HepG2와 Hep3B 세포들은 ${\beta}-lapachone$ 농도 의존적으로 세포의 증식이 억제되었으며 그 형태적 변형도 동반하였다. ${\beta}-lapachone$처리에 의한 암 전이 지표가 되는 IGF-lR, Tjs (ZO-1, claudin-3,-4) 및 Tj 조절인자(${\beta}-catenin$)의 발현을 RT-PCR과 Western blot analysis를 통하여 확인한 결과 ${\beta}-lapachone$ 처리가 IGF-1R의 발현 억제와 Tj 유전자 발현의 증가를 유도함으로써 ${\beta}-lapachone$이 Tj를 강화하여 암세포의 전이 억제작용을 하는 것으로 관찰 되었다. 이상의 결과는 인체 간암세포에서 ${\beta}-lapachone$의 항전이 작용의 이해에 중요한 기초 자료가 될 것으로 생각한다.

다국어 초록 (Multilingual Abstract)

A hallmark of cancers is 'leaky' tight junctions (Tjs). TJs mediated paracellular permeability is elevated and TJs maintained cell polarity is frequently lost. Concomitantly, TJs-associated proteins including members of the claudin family of proteins are dysregulated. Recent findings indicate that these TJs changes can contribute to cancer progression. In this study, we examined the effects of ${\beta}-lapachone$, a quinone compound obtained from the bark of the lapacho tree (Tabebuia avellanedae), on the Tjs-associated regulators in human hepatocarcinoma cell lines, HepG2 and Hep3B. ${\beta}-lapachone$ treatment downregulated the levels of insulin-like growth factor 1 receptor (IGF-lR) proteins in both HepG2 and Hep3B cells. But the levels of claudin-3 and -4 proteins were increased in ${\beta}-lapachone$-treated HepG2 and Hep3B cells. And also the zonnula occludens-l (la-I) and p-catenin protein levels by ${\beta}-lapachone$ were increased in a time-dependent manner. However, claudin-3 and -4 mRNA levels were uninhibited by ${\beta}-lapachone$ in HepG2 and Hep3B. The present results suggest that the upregulation of claudin-3 and -4 protein levels by ${\beta}-lapachone$ occurs by a post-transcriptional mechanism and points to a novel mechanism by ${\beta}-lapachone$.

참고문헌 (Reference)

1 Planchon, S. M., "β-lapachone-induced apoptosis in human prostate cancer cells: involvement of QO1/xip3" 267 : 95-106, 2001

2 Choi, B. T., "β-lapachone- induced apoptosis is associated with activation of caspase-3 and inactivation of NF-κB in human colon cancer HCT-116 cells" 14 : 845-850, 2003

3 Gupta, D., "β-lapachone, a novel plant product, overcomes drug resistance in human multiple myeloma cells" 30 : 711-720, 2002

4 Schaffner-Sabba, K., "β-lapachone synthesis of derivatives and activities in tumor models" 27 : 990-994, 1984

5 Huang, L, "β-lapachone induces cell cycle arrest and apoptosis in human colon cancer cells" 5 : 711-720, 1999

6 Van Itallie, C. M., "The Molecular Physiology of tight junction pores" 19 : 331-338, 2004

7 Choi, Y. H., "Supression of human prostate cancer cell growth by β-lapachone via down-regulation of pRB phosphrylation and induction of Cdk inhibitor p21(WAF1/CIP1)" 36 : 223-229, 2003

8 Choi, Y. H., "Phosphorylation of p53, induction of Bax and activation of caspases during β-lapachone-mediated apoptosis in human prostate epithelial cells" 21 : 1293-1299, 2002

9 Kominsky, S. L., "Loss of the tight junction protein claudin-7 correlates with histological grade in both ductal carcinoma in situ and invasive ductal carcinoma of the breast" 22 : 2021-2033, 2003

10 Hoover, K. B., "Loss of the tight junction MAGUK ZO-1 in breast cancer" 153 : 1767-1773, 1998

11 Chau, Y. P., "Involvement of hydrogen peroxide in topoisomerase inhibitor β-lapachone-induced apoptosis and differentiation in human leukemia cells" 24 : 660-670, 1998

12 Andre, F., "Integrins and E-cadherin cooperate with IGF-I to induce migration of epithelial colonic cells" 83 : 497-505, 1999

13 Li, C. J., "Induction of apoptosis by β-lapachone in human prostate cancer cells" 55 : 3712-3715, 1995

14 Don, M. J., "Induction of CDK inhibitors (p21(WAF1) and p27(Kip1)) and Bak in the β-lapachone-induced apoptosis in human prostate cancer cells" 59 : 784-794, 2001

15 Lopez, J. N., "In vitro and in vivo evaluation of the toxicity of 1,4-naphthoquinone and 1,2-naphthoquinone derivatives against Trypanosoma cruzi" 72 : 523-531, 1978

16 Lopez, T., "Elevated levels of IGF-1 receptor convey invasive and metastatic capability in a mouse model of pancreatic islet tumorigenesis" 1 : 339-353, 2002

17 Guiraud, P., "Comparison of antibacterial and antifungal activities of lapachol and β-lapachone" 60 : 373-374, 1994

18 Tokés, A. M., "Claudin-1, -3 and -4 proteins and mRNA expression in benign and malignant breast lesions: a research study" 7 : R296-R305, 2005

19 Morin, P., "Claudin proteins in human cancer: promising new targets for diagnosis and therapy" 65 : 9603-9606, 2005

20 Tagliarino, C., "Calcium is a key signaling molecule in β-lapachone-mediated cell death" 276 : 19150-19159, 2001

21 Shiah, S. G., "Activation of c-Jun NH2-terminal kinase and subsequent CPP32/Yama during topoisomerase inhibitor β-lapachone-induced apoptosis through an oxidation-dependent pathway" 59 : 391-398, 1999

22 Pink, J. J., "Activation of a cysteine protease in MCF-7 and T47D breast cancer cells during β-lapachone-mediated apoptosis" 255 : 144-155, 2000