- Email: [email protected]
P129. BMP signaling stimulates alveolar bone invasion by melanoma cells
Abstract / Differentiation 80 (2010) S17–S63
P129 BMP signaling stimulates alveolar bone invasion by melanoma cells
M. Shin a, H. Fukushima b, H. Furuta b, K. Aoki c, K. Masud c, K. Ohya c, T. Katagiri a, E. Jimi b
S61
dim stained by integrin a6 and CD71, then abri population 6 /CD71 was regarded as stem cells. As expected, overexpression of PKCd dim bri led to the reduction of a6 /CD71 population, suggesting that PKCd is a negative regulator for maintenance of cancer stem cells. Consistent with this notion, colony forming acitivity was markedly decreased by PKCd. Furthermore, overexpression of PKCd led to remarkable decrease of p63 level in SCC13 cells, suggesting that PKCd regulates cancer stem cells negatively through the modulation of p63.
a
Saitama Medical University, Saitama, Japan Kyushu Dental College, Fukuoka, Japan c Tokyo Medical & Dental University, Tokyo, Japan E-mail address: [email protected] (M. Shin) b
Malignant melanoma in the oral cavity has a much poorer prognosis than its counterpart on the skin. Melanoma in the oral cavity is possible to invade alveolar bone. Although bone morphogenetic proteins have been suggested to be involved in the melanoma invasion, molecular mechanisms are unclear because an animal model in vivo of this event has not been established yet. In the present study, we examined roles of BMP signaling in bone invasion by melanoma cells in vivo and in vitro using mouse melanoma cells. Treatment the melanoma cells with BMP-2 induced changes in cell morphology from prominently stretched to spindle. The rhodamine-conjugated phalloidin stain showed that this morphological change was induced by actin reconstitution. Over-expression of a constitutively active BMP receptor in vitro also induced similar morphological changes and expression of N-Cadherin. A cell line stably expressing a constitutively active BMP receptor, but not an empty vector, showed bone invasion in vivo. These findings suggest that BMP signaling stimulates bone invasion by melanoma cells by inducing changes in cell morphology. doi: 10.1016/j.diff.2010.09.135
P130 Role of PKCd in maintenance of cancer stem cells
Y. Lee a, J.C. Lee b, N.J. Jeong a, S.S. Lee a, C.D. Kim a, J.H. Lee a a
Department of Dermatology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea b Department of Internal Medicine, Daejeon Veterans Hospital, Daejeon, Republic of Korea E-mail address: [email protected] (J.H. Lee)
Cancer stem cells, also called tumor initiating cells, are characterized by their stem-like properties in diverse human cancers. In concordant with a concept of stem cells in normal epithelial cells, squamous cell carcinoma (SCC) cell lines are reported to contain distinct sub-populations with different growth and tumorigenic potential. To investigate the potential role of PKCd in SCC, we first examined the expression of PKCd in human SCC tissues. As compared with normal skins, PKCd expression was generally decreased in SCC tissues. This result suggests that PKCd may have anti-tumorigenic effect in the development of SCC. To investigate the possible role of PKCd in SCC, we overexpressed PKCd in SCC13 cells by recombinant adenovirus. After adenoviral overexpression of PKCd, cell growth of SCC13 was markedly reduced, determined by [3H]thymidine incorporation assay. We hypothesized that PKCd may affect the cancer stemness of SCC13 cells, then determined the change of stem cell population by flow cytometry. Cells were double-
doi: 10.1016/j.diff.2010.09.136
P131 Balanced ubiquitylation and deubiquitylation of Frizzled regulate cellular responsiveness to Wg/Wnt
Akiko Mukai a,b, Miki Yamamoto-Hino a,c, Wakae Awano d, Wakako Watanabe e, Hideyuki Okano c, Masayuki Komada b, Satoshi Goto a,c,d a
Research Group of Glycobiology and Glycotechnology, MitsubishiKagaku Institute of Life Sciences, Machida, Japan b Department of Biological Sciences, Tokyo Institute of Technology, Yokohama, Japan c Advanced Medical Research Laboratory, Research Division, Mitsubishi Tanabe Pharma, Yokohama, Japan d Mutant Flies Laboratory, Mitsubishi-Kagaku Institute of Life Sciences, Machida, Japan e Advanced Medical Research Laboratory, Research Division, Mitsubishi Tanabe Pharma, Yokohama, Japan E-mail address: [email protected] (S. Goto)
Wingless (Wg)/Wnt has been proposed to exert various functions as a morphogen depending on the levels of its signaling. Therefore, not just the concentration of Wg/Wnt, but also the responsiveness of Wg/Wnt target cells to the ligand, must play a crucial role in controlling cellular outputs. Here we show that a balance of ubiquitylation and deubiquitylation of the Wg/Wnt receptor Frizzled determines the cellular responsiveness to Wg/Wnt both in mammalian cells and in Drosophila, and that the cell surface level of Frizzled is regulated by deubiquitylating enzyme UBPY/USP8. While ubiquitylated Frizzled underwent lysosomal trafficking and degradation, UBPY/USP8-dependent deubiquitylation led to recycling of Frizzled to the plasma membrane, thereby elevating its surface level. Importantly, a gain and loss of UBPY/USP8 function led to up- and down-regulation, respectively, of canonical Wg/Wnt signaling. These results unveil a novel mechanism that regulates the cellular responsiveness to Wg/Wnt by controlling the cell surface level of Frizzled. doi: 10.1016/j.diff.2010.09.137
P132 Methylation by protein arginine methyltransferase1 augments stability of Axin
S. Moon, B. Cha, W. Kim, B. Hwang, E. Jho Department of Life Science, University of Seoul, Seoul, Republic of Korea E-mail address: [email protected] (E. Jho)
P129 BMP signaling stimulates alveolar bone invasion by melanoma cells
M. Shin a, H. Fukushima b, H. Furuta b, K. Aoki c, K. Masud c, K. Ohya c, T. Katagiri a, E. Jimi b
S61
dim stained by integrin a6 and CD71, then abri population 6 /CD71 was regarded as stem cells. As expected, overexpression of PKCd dim bri led to the reduction of a6 /CD71 population, suggesting that PKCd is a negative regulator for maintenance of cancer stem cells. Consistent with this notion, colony forming acitivity was markedly decreased by PKCd. Furthermore, overexpression of PKCd led to remarkable decrease of p63 level in SCC13 cells, suggesting that PKCd regulates cancer stem cells negatively through the modulation of p63.
a
Saitama Medical University, Saitama, Japan Kyushu Dental College, Fukuoka, Japan c Tokyo Medical & Dental University, Tokyo, Japan E-mail address: [email protected] (M. Shin) b
Malignant melanoma in the oral cavity has a much poorer prognosis than its counterpart on the skin. Melanoma in the oral cavity is possible to invade alveolar bone. Although bone morphogenetic proteins have been suggested to be involved in the melanoma invasion, molecular mechanisms are unclear because an animal model in vivo of this event has not been established yet. In the present study, we examined roles of BMP signaling in bone invasion by melanoma cells in vivo and in vitro using mouse melanoma cells. Treatment the melanoma cells with BMP-2 induced changes in cell morphology from prominently stretched to spindle. The rhodamine-conjugated phalloidin stain showed that this morphological change was induced by actin reconstitution. Over-expression of a constitutively active BMP receptor in vitro also induced similar morphological changes and expression of N-Cadherin. A cell line stably expressing a constitutively active BMP receptor, but not an empty vector, showed bone invasion in vivo. These findings suggest that BMP signaling stimulates bone invasion by melanoma cells by inducing changes in cell morphology. doi: 10.1016/j.diff.2010.09.135
P130 Role of PKCd in maintenance of cancer stem cells
Y. Lee a, J.C. Lee b, N.J. Jeong a, S.S. Lee a, C.D. Kim a, J.H. Lee a a
Department of Dermatology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea b Department of Internal Medicine, Daejeon Veterans Hospital, Daejeon, Republic of Korea E-mail address: [email protected] (J.H. Lee)
Cancer stem cells, also called tumor initiating cells, are characterized by their stem-like properties in diverse human cancers. In concordant with a concept of stem cells in normal epithelial cells, squamous cell carcinoma (SCC) cell lines are reported to contain distinct sub-populations with different growth and tumorigenic potential. To investigate the potential role of PKCd in SCC, we first examined the expression of PKCd in human SCC tissues. As compared with normal skins, PKCd expression was generally decreased in SCC tissues. This result suggests that PKCd may have anti-tumorigenic effect in the development of SCC. To investigate the possible role of PKCd in SCC, we overexpressed PKCd in SCC13 cells by recombinant adenovirus. After adenoviral overexpression of PKCd, cell growth of SCC13 was markedly reduced, determined by [3H]thymidine incorporation assay. We hypothesized that PKCd may affect the cancer stemness of SCC13 cells, then determined the change of stem cell population by flow cytometry. Cells were double-
doi: 10.1016/j.diff.2010.09.136
P131 Balanced ubiquitylation and deubiquitylation of Frizzled regulate cellular responsiveness to Wg/Wnt
Akiko Mukai a,b, Miki Yamamoto-Hino a,c, Wakae Awano d, Wakako Watanabe e, Hideyuki Okano c, Masayuki Komada b, Satoshi Goto a,c,d a
Research Group of Glycobiology and Glycotechnology, MitsubishiKagaku Institute of Life Sciences, Machida, Japan b Department of Biological Sciences, Tokyo Institute of Technology, Yokohama, Japan c Advanced Medical Research Laboratory, Research Division, Mitsubishi Tanabe Pharma, Yokohama, Japan d Mutant Flies Laboratory, Mitsubishi-Kagaku Institute of Life Sciences, Machida, Japan e Advanced Medical Research Laboratory, Research Division, Mitsubishi Tanabe Pharma, Yokohama, Japan E-mail address: [email protected] (S. Goto)
Wingless (Wg)/Wnt has been proposed to exert various functions as a morphogen depending on the levels of its signaling. Therefore, not just the concentration of Wg/Wnt, but also the responsiveness of Wg/Wnt target cells to the ligand, must play a crucial role in controlling cellular outputs. Here we show that a balance of ubiquitylation and deubiquitylation of the Wg/Wnt receptor Frizzled determines the cellular responsiveness to Wg/Wnt both in mammalian cells and in Drosophila, and that the cell surface level of Frizzled is regulated by deubiquitylating enzyme UBPY/USP8. While ubiquitylated Frizzled underwent lysosomal trafficking and degradation, UBPY/USP8-dependent deubiquitylation led to recycling of Frizzled to the plasma membrane, thereby elevating its surface level. Importantly, a gain and loss of UBPY/USP8 function led to up- and down-regulation, respectively, of canonical Wg/Wnt signaling. These results unveil a novel mechanism that regulates the cellular responsiveness to Wg/Wnt by controlling the cell surface level of Frizzled. doi: 10.1016/j.diff.2010.09.137
P132 Methylation by protein arginine methyltransferase1 augments stability of Axin
S. Moon, B. Cha, W. Kim, B. Hwang, E. Jho Department of Life Science, University of Seoul, Seoul, Republic of Korea E-mail address: [email protected] (E. Jho)