• Asian Pacific Journal of Cancer Prevention
• /
• 제17권3호
• /
• pp.957-963
• /
• 2016

Cellular senescence, a barrier to tumorigenesis, controls aberrant proliferation of cells. We here aimed to investigate cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines using five different inducing agents: 5-aza-2'deoxycytidine, bromodeoxyuridine, interferons ($IFN{\beta}$ and $IFN{\gamma}$), and hydrogen peroxide. We analyzed senescence characteristics, colony formation ability, expression of genes involved in cell cycling and interferon signaling pathways, and protein levels. Treatment with all five agents decreased cell proliferation and induced cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines with different degrees of growth-inhibitory effects depending on cell type and origin. Bromodeoxyuridine gave the strongest stimulus to inhibit growth and induce senescence in most cell lines tested. Expression of p21 and interferon related genes was upregulated in most conditions. The fact that bromodeoxyuridine had the strongest effects on growth inhibition and senescence induction implies that senescence in cholangiocarcinoma cells is likely controlled by DNA damage response pathways relating to the p53/p21 signaling. In addition, interferon signaling pathways may partly regulate this mechanism in cholangiocarcinoma cells.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권4호
• /
• pp.2503-2508
• /
• 2013

Histone deacetylation mediated by histone deacetylases (HDACs) has been reported as one of the epigenetic mechanisms associated with tumorigenesis. The poor responsiveness of anticancer drugs found with cholangiocarcinoma (CCA) leads to short survival rate. We aimed to investigate mRNA expression of HDACs class I and II, and the effect of HDAC inhibitors, suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA), in CCA in vitro. Expression of HDACs was studied in CCA cell lines (M213, M214 and KKU-100) and an immortal cholangiocyte (MMNK1) by semi-quantitative reverse transcription-PCR. SAHA and VPA, as well as a classical chemotherapeutic drug 5 -fluorouacil (5-FU) were used in this study. Cell proliferation was determined by sulforhodamine assay. $IC_{50}$ and $IC_{20}$ were then analyzed for each agent and cell line. Moreover, synergistic potentional of VPA or SAHA in combination with 5-FU at sub toxic does ($IC_{20}$) of each agent was also evaluated. Statistic difference of HDACs expression or cell proliferation in each experimental condition was analyzed by Student's t-test. The result demonstrated that HDACs were expressed in all studied cell types. Both SAHA and VPA inhibited cell proliferation in a dose-dependent manner. Interestingly, KKU-100 which was less senstitive to classical chemotheraoeutic 5-FU was highly was sensitive to HDAC inhibitors. Simultaneous combination of subtoxic doses of HDAC inhibitors and 5-FU signiicantly inhibited cell proliferation in CCA cell lines compared to single sgent treatment($P{\leq}0.01$), while sequentially combined treatments were less effective. The present study showed inhibitory effects of HDACIs on cell proliferation in CCA cell lines, with synergistic antitumor potential demonstrated by simultaneous combination of VPA or SAHA with 5-FU, suggesting a novel alternative therapeutic strategy in effective treatment of CCA.

• 생명과학회지
• /
• 제21권12호
• /
• pp.1772-1777
• /
• 2011

본 연구는 흰쥐의 담관 세포와 간세포에서 CFTR과 $AE1{\cdot}AE2{\cdot}AE3$ 유전자들의 발현 유무를 조사하고 흰쥐에서 담관 결찰 후 AE2 유전자의 발현의 변화를 관찰하고자 하였다. 200-250 g의 Sprague Dawley 계 흰쥐 24마리의 총담관을 결찰한 후 4 주 동안 1 주일에 6마리씩 희생하여 간세포와 담관 세포를 분리하였다. 6마리는 대조군으로 사용하여 간세포와 담관 세포에서 CFTR 유전자와 $AE1{\cdot}AE2$와 AE3 유전자 발현을 조사하고 담관 결찰 후 1 2 3 4주 간격으로 AE2 유전자 발현을 조사하였다. $AE1{\cdot}AE2$ 와 AE3는 간세포와 담관 세포에서 발현되었고 CFTR은 담관 세포에서만 발현되었다. 담관 결찰 담관세포군에서 AE2 유전자의 발현은 대조군인 정상 담관세포군에 비해서 낮았다. 결찰 담관세포군에서 AE2 유전자의 발현은 결찰 기간에 따라 차이가 없었다. 담관 결찰 간세포군에서 AE2 유전자의 발현은 대조군인 정상 간세포군에 비해서 경계적 유의성을 보이며 증가하는 경향이 있었다. 결찰 간세포군에서 AE2 유전자의 발현은 결찰 기간에 따라 차이는 없었다. 따라서 $CFTR{\cdot}AE1{\cdot}AE2$ 그리고AE3 는 간세포와 담관 세포에서 중탄산염이온과 수액을 매개하는 주된 이온 전달체라는 사실을 고려할 때 담도 담즙정체 간질환에서CFTR과 AE2 발현의 변화는 병리학적 기전에 중요한 역할을 할 수 있으리라고 생각된다.