Influence of L-Arginine on Cell Proliferation, Inducible Nitric Oxide Synthase Expression and Cell Cycle of Human Colon Carcinoma Cell Line LS174 Through Nitric Oxide Pathway_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

HUO Xiongwei ¹ , GAO Yanfeng ² , MA Qingyong ¹ , SUN Xuejun ¹

1.Department of General Surgery, The First Affiliated Hospital, Medical School of Xi’an Jiaotong University, Xi’an 710061, China;;
2. Department of Anesthesiology, The First Affiliated Hospital, Medical School of Xi’an Jiaotong University, Xi’an 710061, China;

Corresponding author：

Keywords：

L-arginine; Nitric oxide; Inducible nitric oxide synthase; Cell cycle; Human colon carcinoma cell line LS174; Cell proliferation

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Objective To study the effects of L-arginine （L-Arg） on cell proliferation, inducible nitric oxide synthase (iNOS) expression and cell cycle in human colon carcinoma cell line LS174 through nitric oxide (NO) pathway. Methods LS174 cells were cultured in medium with L-Arg at different concentrations for different times. MTT method was employed to evaluate the level of the cell proliferation. The production of NO in culture supernatants of LS174 cell was detected with enzyme reduction of nitrate. The distribution of the cell cycle was detected with the flow cytometry (FCM). The expression level of iNOS in the cells was determined by Western blot and SP immunocytochemical staining method. Results The growth of LS174 was promoted by the L-Arg at low concentration (0.125 mmol/L) and inhibited at high concentrations (0.5， 2， 8 and 32 mmol/L). The level of NO was increased with the increasing concentration of L-Arg in culture medium. To compare with the control group, the ratio of cells at S phase was increased after 48 hours’ treatments with high concentrations (0.5, 2, 8 and 32 mmol/L) of L-Arg (P＜0.05, P＜0.01); while there was no obvious difference after treatments with low concentration (0.125 mmol/L) of L-Arg (P gt;0.05). With the increase of the concentration of L-Arg, the expression of iNOS was increased as compared with control group. The higher the concentration of L-Arg was, the better the effect. Conclusion L-Arg can induce the expression of iNOS resulting in increase the production of nitric oxide (NO). Low concentration of L-Arg can promote the growth of LS174 cells, while high concentration ones can inhibit growth and proliferation. The high concentration of L-Arg could induce S phase arrestion in the cell cycle.

Citation： HUO Xiongwei,GAO Yanfeng,MA Qingyong,SUN Xuejun. Influence of L-Arginine on Cell Proliferation, Inducible Nitric Oxide Synthase Expression and Cell Cycle of Human Colon Carcinoma Cell Line LS174 Through Nitric Oxide Pathway. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(11): 880-885. doi: Copy

1.	Mori M, Gotoh T. Arginine metabolic enzymes, nitric oxide and infection [J]. J Nutr, 2004; 134(10 Suppl): 2820S-2825S.
2.	Keklikoglu N, Koray M, Kocaelli H, et al. iNOS expression in oral and gastrointestinal tractmucosa [J]. Dig Dis Sci, 2008; 53(6): 1437-1442.
3.	Kotamraju S, Williams CL, Kalyanaraman B. Statin-induced breast cancer cell death: role of inducible nitric oxide and arginase-dependent pathways [J]. Cancer Res, 2007; 67 (15): 7386-7394.
4.	Vissersy YL, Dejong CH, Luiking YC, et al. Plasma arginine concentrations are reduced in cancer patients: evidence for arginine deficiency? [J]. Am J Clin Nutr, 2005; 81 (5): 1142-1146.
5.	Caso G, McNurlan MA, McMillan ND, et al. Tumour cell growth in culture: dependence on arginine [J]. Clin Sci (Lond), 2004; 107(4): 371-379.
6.	Ma Q, Wang Y, Gao X, et al. L-arginine reduces cell proliferation and ornithine decarboxylase activity in patients with colorectal adenoma and adenocarcinoma [J]. Clin Cancer Res, 2007; 13(24): 7407-7412.
7.	Lin Y, Wang LN, Xi YH, et al. L-arginine inhibits isoproterenol-induced cardiac hypertrophy through nitric oxide and polyamine pathways [J]. Basic Clin Pharmacol Toxicol, 2008; 103(2): 124-130.
8.	Bracci M, Tomasetti M, Malavolta M, et al. L-arginine reduces mercury accumulation in thymus of mercury-exposed mice: role of nitric oxide synthase activity and metallothioneins [J]. Ind Health, 2008; 46(6): 567-574.
9.	Schwedhelm E, Maas R, Freese R, et al. Pharmacokinetic and pharmacodynamic properties of oral L-citrulline and L-arginine: impact on nitric oxide metabolism [J]. Br J Clin Pharmacol, 2008; 65(1): 51-59.
10.	王广义，王旭. 人胃癌VEGF 和NOS 的表达与肿瘤血管生成的关系 [J]. 中国普外基础与临床杂志, 2004; 11(1)∶ 55-57.
11.	张剑, 何生, 李茂德. 肝细胞癌中iNOS 和p53 蛋白的表达及其与肿瘤血管形成的关系 [J]. 中国普外基础与临床杂志, 2004; 11(5)： 396-398.
12.	Gardner PR, Gardner AM, Brashear WT, et al. Hemoglobins dioxygenate nitric oxide with high fidelity [J]. J Inorg Biochem, 2006; 100(4): 542-550.
13.	Chen HJ, Hsieh CJ, Shen LC, et al. Characterization of DNA-protein cross-links induced by oxanine: cellular damage derived from nitric oxide and nitrous acid [J]. Biochemistry, 2007; 46(13): 3952-3965.
14.	Zhao X, Mohaupt M, Jiang J, et al. Tumor necrosis factor receptor 2-mediated tumor suppression is nitric oxide dependent and involves angiostasis [J]. Cancer Res, 2007; 67(9): 4443-4450.
15.	Yung HW, Bal-Price AK, Brown GC, et al. Nitric oxide-induced cell death of cerebrocortical murine astrocytes is mediated through p53-and Bax-dependent pathways [J]. J Neurochem, 2004; 89(4): 812-821.
16.	Stotz WH, Li D, Johns RA. Exogenous nitric oxide upregulates p21waf1/cip1 in pulmonary microvascular smooth muscle cells [J]. J Vasc Res, 2004; 41(3): 211-219.
17.	Frederiksen LJ, Sullivan R, Maxwell LR, et al. Chemosensitization of cancer in vitro and in vivo by nitric oxide signaling [J]. Clin Cancer Res, 2007; 13(7): 2199-2206.
18.	Moulik S, Chatterjee A. Vascular endothelial growth factor (VEGF) & tumour angiogenesis [J]. Indian J Med Res, 2007; 125(6): 747-751.
19.	Shigyo H, Nonaka S, Katada A, et al. Inducible nitric oxide synthase expression in various l aryngeal lesions in relation to carcinogenesis, angiogenesis, and patients prognosis [J]. Acta Otolaryngol, 2007; 127(9): 970-979.
20.	Kolb JP. Mechanisms involved in the pro-and anti-apoptotic role of NO in human leukemia [J]. Leukemia, 2000; 14(9): 1685-1694.
21.	Hammadi A, Billard C, Faussat AM, et al. Stimulation of iNOS expression and apoptosis resistance in B-cell chronic lymphocytic leukemia (B-CLL) cells through engagement of Toll-like receptor 7 (TLR-7) and NF-kappaB activation [J]. Nitric Oxide, 2008; 19(2): 138-145.
22.	Evan GI, Vousden KH. Proliferation, cell cycle and apoptosis in cance [J]. Nature, 2001; 411(6835): 342-348.
23.	Massagué J. G1 cell-cycle control and cancer [J]. Nature, 2004; 432(7015): 298-306.
24.	Jiang W, Zhu Z, Bhatia N, et al. Mechanisms of energy restriction: effects of corticosterone on cell growth, cell cycle machinery, and apoptosis [J]. Cancer Res, 2002; 62(18): 5280-5287.
25.	Sarkar R, Gordon D, Stanley JC, et al. Cell cycle effects of nitric oxide on vascular smooth muscle cells [J]. Am J Physiol Heart Circ Physiol, 1997; 272(4pt2): H1810-1818.

1. Mori M, Gotoh T. Arginine metabolic enzymes, nitric oxide and infection [J]. J Nutr, 2004; 134(10 Suppl): 2820S-2825S.
2. Keklikoglu N, Koray M, Kocaelli H, et al. iNOS expression in oral and gastrointestinal tractmucosa [J]. Dig Dis Sci, 2008; 53(6): 1437-1442.
3. Kotamraju S, Williams CL, Kalyanaraman B. Statin-induced breast cancer cell death: role of inducible nitric oxide and arginase-dependent pathways [J]. Cancer Res, 2007; 67 (15): 7386-7394.
4. Vissersy YL, Dejong CH, Luiking YC, et al. Plasma arginine concentrations are reduced in cancer patients: evidence for arginine deficiency? [J]. Am J Clin Nutr, 2005; 81 (5): 1142-1146.
5. Caso G, McNurlan MA, McMillan ND, et al. Tumour cell growth in culture: dependence on arginine [J]. Clin Sci (Lond), 2004; 107(4): 371-379.
6. Ma Q, Wang Y, Gao X, et al. L-arginine reduces cell proliferation and ornithine decarboxylase activity in patients with colorectal adenoma and adenocarcinoma [J]. Clin Cancer Res, 2007; 13(24): 7407-7412.
7. Lin Y, Wang LN, Xi YH, et al. L-arginine inhibits isoproterenol-induced cardiac hypertrophy through nitric oxide and polyamine pathways [J]. Basic Clin Pharmacol Toxicol, 2008; 103(2): 124-130.
8. Bracci M, Tomasetti M, Malavolta M, et al. L-arginine reduces mercury accumulation in thymus of mercury-exposed mice: role of nitric oxide synthase activity and metallothioneins [J]. Ind Health, 2008; 46(6): 567-574.
9. Schwedhelm E, Maas R, Freese R, et al. Pharmacokinetic and pharmacodynamic properties of oral L-citrulline and L-arginine: impact on nitric oxide metabolism [J]. Br J Clin Pharmacol, 2008; 65(1): 51-59.
10. 王广义，王旭. 人胃癌VEGF 和NOS 的表达与肿瘤血管生成的关系 [J]. 中国普外基础与临床杂志, 2004; 11(1)∶ 55-57.
11. 张剑, 何生, 李茂德. 肝细胞癌中iNOS 和p53 蛋白的表达及其与肿瘤血管形成的关系 [J]. 中国普外基础与临床杂志, 2004; 11(5)： 396-398.
12. Gardner PR, Gardner AM, Brashear WT, et al. Hemoglobins dioxygenate nitric oxide with high fidelity [J]. J Inorg Biochem, 2006; 100(4): 542-550.
13. Chen HJ, Hsieh CJ, Shen LC, et al. Characterization of DNA-protein cross-links induced by oxanine: cellular damage derived from nitric oxide and nitrous acid [J]. Biochemistry, 2007; 46(13): 3952-3965.
14. Zhao X, Mohaupt M, Jiang J, et al. Tumor necrosis factor receptor 2-mediated tumor suppression is nitric oxide dependent and involves angiostasis [J]. Cancer Res, 2007; 67(9): 4443-4450.
15. Yung HW, Bal-Price AK, Brown GC, et al. Nitric oxide-induced cell death of cerebrocortical murine astrocytes is mediated through p53-and Bax-dependent pathways [J]. J Neurochem, 2004; 89(4): 812-821.
16. Stotz WH, Li D, Johns RA. Exogenous nitric oxide upregulates p21waf1/cip1 in pulmonary microvascular smooth muscle cells [J]. J Vasc Res, 2004; 41(3): 211-219.
17. Frederiksen LJ, Sullivan R, Maxwell LR, et al. Chemosensitization of cancer in vitro and in vivo by nitric oxide signaling [J]. Clin Cancer Res, 2007; 13(7): 2199-2206.
18. Moulik S, Chatterjee A. Vascular endothelial growth factor (VEGF) & tumour angiogenesis [J]. Indian J Med Res, 2007; 125(6): 747-751.
19. Shigyo H, Nonaka S, Katada A, et al. Inducible nitric oxide synthase expression in various l aryngeal lesions in relation to carcinogenesis, angiogenesis, and patients prognosis [J]. Acta Otolaryngol, 2007; 127(9): 970-979.
20. Kolb JP. Mechanisms involved in the pro-and anti-apoptotic role of NO in human leukemia [J]. Leukemia, 2000; 14(9): 1685-1694.
21. Hammadi A, Billard C, Faussat AM, et al. Stimulation of iNOS expression and apoptosis resistance in B-cell chronic lymphocytic leukemia (B-CLL) cells through engagement of Toll-like receptor 7 (TLR-7) and NF-kappaB activation [J]. Nitric Oxide, 2008; 19(2): 138-145.
22. Evan GI, Vousden KH. Proliferation, cell cycle and apoptosis in cance [J]. Nature, 2001; 411(6835): 342-348.
23. Massagué J. G1 cell-cycle control and cancer [J]. Nature, 2004; 432(7015): 298-306.
24. Jiang W, Zhu Z, Bhatia N, et al. Mechanisms of energy restriction: effects of corticosterone on cell growth, cell cycle machinery, and apoptosis [J]. Cancer Res, 2002; 62(18): 5280-5287.
25. Sarkar R, Gordon D, Stanley JC, et al. Cell cycle effects of nitric oxide on vascular smooth muscle cells [J]. Am J Physiol Heart Circ Physiol, 1997; 272(4pt2): H1810-1818.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Influence of L-Arginine on Cell Proliferation, Inducible Nitric Oxide Synthase Expression and Cell Cycle of Human Colon Carcinoma Cell Line LS174 Through Nitric Oxide Pathway

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