T Jacob1, N Clouden, A Hingorani, E Ascher. 1. Department of Surgery, Division of Vascular Surgery, Maimonides Medical Center, Brooklyn New York, New York, NY, USA. tjacob@maimonidesmed.org
Abstract
AIM: Cotinine, the main stable metabolite of nicotine, has been shown to have a biological half-life approximately 10 times longer than nicotine. It has also been demonstrated to have a powerful effect on vascular smooth muscle cell (VSMC) proliferation. Telomerase activation is known to play an important role in cell viability and proliferation. The purpose of our experiment was to evaluate the effect of cotinine on proliferative potential of vascular smooth muscle cells via its effects on telomerase activity. METHODS: Primary cultures of human VSMC obtained from greater saphenous veins were used in this experiment from 3(rd) to 5(th) passage. Cotinine was added in doses equivalent to plasma levels of cotinine in an active smoker by dissolving, 0.0, 2.88x10(-6), 5.76x10(-6), and 1.44x10(-5) mol/L of cotinine in the media. The number of viable cells was assessed by trypan blue exclusion. The Telomeric Repeat Amplification Protocol (TRAP) was used to detect telomerase activity. TRAP products were detected by ELISA. RESULTS: The mitogenic effect of cotinine in VSMC was observed at 48 hours after treatment. The viable cell numbers were significantly increased (4.0x10(7)) at lower doses of cotinine exposure as compared to untreated cultures (2.5x10(5)). At the concentration of 1.44x10(-5) mol/L, cotinine was cytotoxic to VSMCs. Telomerase activity was detected in all sets of VSMC cultures treated with cotinine (P<0.01). CONCLUSIONS: Cotinine causes abnormal cell proliferation as demonstrated by increased cell numbers and reactivation of telomerase in a dose dependent manner. This study demonstrated cotinine's stimulatory effect on human SMC proliferation in vitro at low doses while high doses of cotinine had a toxic effect. These data correlate with the results of other studies concerning the mitogenic effect of cotinine and telomerase activation during cellular proliferative response.
AIM: Cotinine, the main stable metabolite of nicotine, has been shown to have a biological half-life approximately 10 times longer than nicotine. It has also been demonstrated to have a powerful effect on vascular smooth muscle cell (VSMC) proliferation. Telomerase activation is known to play an important role in cell viability and proliferation. The purpose of our experiment was to evaluate the effect of cotinine on proliferative potential of vascular smooth muscle cells via its effects on telomerase activity. METHODS: Primary cultures of humanVSMC obtained from greater saphenous veins were used in this experiment from 3(rd) to 5(th) passage. Cotinine was added in doses equivalent to plasma levels of cotinine in an active smoker by dissolving, 0.0, 2.88x10(-6), 5.76x10(-6), and 1.44x10(-5) mol/L of cotinine in the media. The number of viable cells was assessed by trypan blue exclusion. The Telomeric Repeat Amplification Protocol (TRAP) was used to detect telomerase activity. TRAP products were detected by ELISA. RESULTS: The mitogenic effect of cotinine in VSMC was observed at 48 hours after treatment. The viable cell numbers were significantly increased (4.0x10(7)) at lower doses of cotinine exposure as compared to untreated cultures (2.5x10(5)). At the concentration of 1.44x10(-5) mol/L, cotinine was cytotoxic to VSMCs. Telomerase activity was detected in all sets of VSMC cultures treated with cotinine (P<0.01). CONCLUSIONS:Cotinine causes abnormal cell proliferation as demonstrated by increased cell numbers and reactivation of telomerase in a dose dependent manner. This study demonstrated cotinine's stimulatory effect on human SMC proliferation in vitro at low doses while high doses of cotinine had a toxic effect. These data correlate with the results of other studies concerning the mitogenic effect of cotinine and telomerase activation during cellular proliferative response.
Authors: William H Goodson; Leroy Lowe; David O Carpenter; Michael Gilbertson; Abdul Manaf Ali; Adela Lopez de Cerain Salsamendi; Ahmed Lasfar; Amancio Carnero; Amaya Azqueta; Amedeo Amedei; Amelia K Charles; Andrew R Collins; Andrew Ward; Anna C Salzberg; Annamaria Colacci; Ann-Karin Olsen; Arthur Berg; Barry J Barclay; Binhua P Zhou; Carmen Blanco-Aparicio; Carolyn J Baglole; Chenfang Dong; Chiara Mondello; Chia-Wen Hsu; Christian C Naus; Clement Yedjou; Colleen S Curran; Dale W Laird; Daniel C Koch; Danielle J Carlin; Dean W Felsher; Debasish Roy; Dustin G Brown; Edward Ratovitski; Elizabeth P Ryan; Emanuela Corsini; Emilio Rojas; Eun-Yi Moon; Ezio Laconi; Fabio Marongiu; Fahd Al-Mulla; Ferdinando Chiaradonna; Firouz Darroudi; Francis L Martin; Frederik J Van Schooten; Gary S Goldberg; Gerard Wagemaker; Gladys N Nangami; Gloria M Calaf; Graeme Williams; Gregory T Wolf; Gudrun Koppen; Gunnar Brunborg; H Kim Lyerly; Harini Krishnan; Hasiah Ab Hamid; Hemad Yasaei; Hideko Sone; Hiroshi Kondoh; Hosni K Salem; Hsue-Yin Hsu; Hyun Ho Park; Igor Koturbash; Isabelle R Miousse; A Ivana Scovassi; James E Klaunig; Jan Vondráček; Jayadev Raju; Jesse Roman; John Pierce Wise; Jonathan R Whitfield; Jordan Woodrick; Joseph A Christopher; Josiah Ochieng; Juan Fernando Martinez-Leal; Judith Weisz; Julia Kravchenko; Jun Sun; Kalan R Prudhomme; Kannan Badri Narayanan; Karine A Cohen-Solal; Kim Moorwood; Laetitia Gonzalez; Laura Soucek; Le Jian; Leandro S D'Abronzo; Liang-Tzung Lin; Lin Li; Linda Gulliver; Lisa J McCawley; Lorenzo Memeo; Louis Vermeulen; Luc Leyns; Luoping Zhang; Mahara Valverde; Mahin Khatami; Maria Fiammetta Romano; Marion Chapellier; Marc A Williams; Mark Wade; Masoud H Manjili; Matilde E Lleonart; Menghang Xia; Michael J Gonzalez; Michalis V Karamouzis; Micheline Kirsch-Volders; Monica Vaccari; Nancy B Kuemmerle; Neetu Singh; Nichola Cruickshanks; Nicole Kleinstreuer; Nik van Larebeke; Nuzhat Ahmed; Olugbemiga Ogunkua; P K Krishnakumar; Pankaj Vadgama; Paola A Marignani; Paramita M Ghosh; Patricia Ostrosky-Wegman; Patricia A Thompson; Paul Dent; Petr Heneberg; Philippa Darbre; Po Sing Leung; Pratima Nangia-Makker; Qiang Shawn Cheng; R Brooks Robey; Rabeah Al-Temaimi; Rabindra Roy; Rafaela Andrade-Vieira; Ranjeet K Sinha; Rekha Mehta; Renza Vento; Riccardo Di Fiore; Richard Ponce-Cusi; Rita Dornetshuber-Fleiss; Rita Nahta; Robert C Castellino; Roberta Palorini; Roslida Abd Hamid; Sabine A S Langie; Sakina E Eltom; Samira A Brooks; Sandra Ryeom; Sandra S Wise; Sarah N Bay; Shelley A Harris; Silvana Papagerakis; Simona Romano; Sofia Pavanello; Staffan Eriksson; Stefano Forte; Stephanie C Casey; Sudjit Luanpitpong; Tae-Jin Lee; Takemi Otsuki; Tao Chen; Thierry Massfelder; Thomas Sanderson; Tiziana Guarnieri; Tove Hultman; Valérian Dormoy; Valerie Odero-Marah; Venkata Sabbisetti; Veronique Maguer-Satta; W Kimryn Rathmell; Wilhelm Engström; William K Decker; William H Bisson; Yon Rojanasakul; Yunus Luqmani; Zhenbang Chen; Zhiwei Hu Journal: Carcinogenesis Date: 2015-06 Impact factor: 4.944
Authors: Amancio Carnero; Carmen Blanco-Aparicio; Hiroshi Kondoh; Matilde E Lleonart; Juan Fernando Martinez-Leal; Chiara Mondello; A Ivana Scovassi; William H Bisson; Amedeo Amedei; Rabindra Roy; Jordan Woodrick; Annamaria Colacci; Monica Vaccari; Jayadev Raju; Fahd Al-Mulla; Rabeah Al-Temaimi; Hosni K Salem; Lorenzo Memeo; Stefano Forte; Neetu Singh; Roslida A Hamid; Elizabeth P Ryan; Dustin G Brown; John Pierce Wise; Sandra S Wise; Hemad Yasaei Journal: Carcinogenesis Date: 2015-06 Impact factor: 4.944