A novel protein derived from lamprey supraneural body tissue with efficient cytocidal actions against tumor cells

Cell Communication and Signaling, Oct 2017

In previous research, we found that cell secretion from the adult lamprey supraneural body tissues possesses cytocidal activity against tumor cells, but the protein with cytocidal activity was unidentified. A novel lamprey immune protein (LIP) as defense molecule was first purified and identified in jawless vertebrates (cyclostomes) using hydroxyapatite column and Q Sepharose Fast Flow column. After LIP stimulation, morphological changes of tumor cells were analysed and measured whether in vivo or in vitro. LIP induces remarkable morphological changes in tumor cells, including cell blebbing, cytoskeletal alterations, mitochondrial fragmentation and endoplasmic reticulum vacuolation, and most of the cytoplasmic and organelle proteins are released following treatment with LIP. LIP evokes an elevation of intracellular calcium and inflammatory molecule levels. Our analysis of the cytotoxic mechanism suggests that LIP can upregulate the expression of caspase 1, RIPK1, RIP3 to trigger pyroptosis and necroptosis. To examine the effect of LIP in vivo, tumor xenograft experiments were performed, and the results indicated that LIP inhibits tumor growth without damage to mice. In addition, the cytotoxic action of LIP depended on the phosphatidylserine (PS) content of the cell membrane. These observations suggest that LIP plays a crucial role in tumor cell survival and growth. The findings will also help to elucidate the mechanisms of host defense in lamprey.

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A novel protein derived from lamprey supraneural body tissue with efficient cytocidal actions against tumor cells

Pang et al. Cell Communication and Signaling A novel protein derived from lamprey supraneural body tissue with efficient cytocidal actions against tumor cells Yue Pang 0 1 Changzhi Li 0 1 Shiyue Wang 0 1 Wei Ba 0 1 Tao Yu 0 1 Guangying Pei 0 1 Dan Bi 0 1 Hongfang Liang 0 1 Xiong Pan 0 1 Ting Zhu 0 1 Meng Gou 0 1 Yinglun Han 0 1 Qingwei Li 0 1 0 Lamprey Research Center, Liaoning Normal University , Dalian 116081 , China 1 College of Life Science, Liaoning Normal University , Dalian 116081 , China Background: In previous research, we found that cell secretion from the adult lamprey supraneural body tissues possesses cytocidal activity against tumor cells, but the protein with cytocidal activity was unidentified. Methods: A novel lamprey immune protein (LIP) as defense molecule was first purified and identified in jawless vertebrates (cyclostomes) using hydroxyapatite column and Q Sepharose Fast Flow column. After LIP stimulation, morphological changes of tumor cells were analysed and measured whether in vivo or in vitro. Results: LIP induces remarkable morphological changes in tumor cells, including cell blebbing, cytoskeletal alterations, mitochondrial fragmentation and endoplasmic reticulum vacuolation, and most of the cytoplasmic and organelle proteins are released following treatment with LIP. LIP evokes an elevation of intracellular calcium and inflammatory molecule levels. Our analysis of the cytotoxic mechanism suggests that LIP can upregulate the expression of caspase 1, RIPK1, RIP3 to trigger pyroptosis and necroptosis. To examine the effect of LIP in vivo, tumor xenograft experiments were performed, and the results indicated that LIP inhibits tumor growth without damage to mice. In addition, the cytotoxic action of LIP depended on the phosphatidylserine (PS) content of the cell membrane. Conclusions: These observations suggest that LIP plays a crucial role in tumor cell survival and growth. The findings will also help to elucidate the mechanisms of host defense in lamprey. Lamprey; LIP; Cytotoxic activity; Inflammatory; Phosphatidylserine Background The adaptive immune system of jawed vertebrates can remove exogenous pathogens or abnormal cells, primarily by cytotoxicity against target cells. In addition to antibodydependent cellular cytotoxicity and the antibody-mediated classical complement pathway, cytotoxic T lymphocytes (CTLs) are able to kill target cells infected by viruses [ 1 ]. The ability of CTLs to kill target cells primarily depends on the amount of cytoplasmic granules, such as perforin and granzyme. They induce programmed cell death in the target cell, thus eliminating cells that have become cancerous or are infected with viruses or bacteria [ 2 ]. In previous research, we observed that lamprey antisera displayed complement dependent cytolytic effects against bacteria and tumor cells [ 3 ]. In this study, a novel protein was discovered in the supraneural body (SB) of adult lamprey that has cytotoxic activity against human MCF-7 and K562 cells. We obtained the gene encoding this novel protein, called lamprey immune protein (LIP), and examined the cytotoxic activities of LIP against a variety of cultured human cells. LIP was found to have strong cytocidal effects against various human cells in vitro and in vivo. In addition, LIP specifically bound to the plasma membrane of tumor cells and then rapidly increased the membrane permeability and dramatically altered the organelle morphologies. Our analyses of the cytocidal effects of this protein demonstrate the following: (i) LIP increases the membrane permeability and alters the organelle morphologies; (ii) evokes an elevation of intracellular calcium, and inflammatory molecule levels; (iii) LIP preferentially kills tumor cells in vitro and in vivo without damage to mice; (iv) LIP upregulates the expression of caspase 1, RIPK1, RIP3 to trigger pyroptosis and necroptosis, and (v) LIP specifically recognizes and binds to PS on the plasma membrane of tumor cells. Thus, LIP is a cell-discriminating and membrane-targeting protein that causes irreversible intracellular decay in tumor cells. Because LIP possesses highly selective cytotoxicity toward human cells and has the potential to specifically recognize and kill some classes of tumor cells, the possibility of its application in medical and biological fields is anticipated. Methods Animals and cell culture Adult lampreys (Lethenteron camtschaticum) weighing 121-152 g were obtained in December 2015 from the Tongjiang Valley of Songhua River, Heilongjiang Province, China. These lampreys were kept at 10 °C in glass tanks with recirculating fresh water at Liaoning Normal University. The animal experiments were performed in accordance with the regulations of the Animal Welfare and Research Ethics Committee of the Institute of Dalian Medical University’s Animal Care protocol (Permit Number: SCXK2008-0002). Human cells used, breast adenocarcinoma cell MCF-7, hepatocyte cancer HepG2, c (...truncated)


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Yue Pang, Changzhi Li, Shiyue Wang, Wei Ba, Tao Yu, Guangying Pei, Dan Bi, Hongfang Liang, Xiong Pan, Ting Zhu, Meng Gou, Yinglun Han, Qingwei Li. A novel protein derived from lamprey supraneural body tissue with efficient cytocidal actions against tumor cells, Cell Communication and Signaling, 2017, pp. 42, DOI: 10.1186/s12964-017-0198-6