Citrus Reticulata Blanco Induces Apoptosis in Human Gastric Cancer Cells SNU-668

Abstract: Citrus fruits have been known to reduce the proliferation of many cancer cells. The antiproliferative effects of Citrus reticulata Blanco (CR) extract, the immature tangerine peel, on human gastric cancer cell line SNU-668 were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6-diamidineo-2-phenylindole staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, reverse transcription-polymerase chain reaction expressions of BCL-2, BAX and CASP-3 genes, caspase-3 activity, and immunocytochemistry of caspase-3. From the results of the morphological and biochemical assays, CR (50 μg/ml) increased the apoptosis of human gastric cancer cells with typical apoptotic characteristics, including morphological changes of chromatin condensation and apoptotic body formation. CR (50 μg/ml) reduced the expression of BCL-2, whereas the expression of BAX and CASP-3 was increased compared with the control group. Furthermore, caspase-3 activity and caspase-3 protein expression in the CR-treated group was significantly increased compared with that in control group. These results suggest that CR may induce the apoptosis through the caspase-3 pathway in human gastric cancer cells.

[1]  K. Okumura,et al.  Apoptosis Induced by the Flavonoid from Lemon Fruit (Citrus limon BURM. f.) and Its Metabolites in HL-60 Cells , 2000, Bioscience, biotechnology, and biochemistry.

[2]  J. Manthey,et al.  Antiproliferative activities of citrus flavonoids against six human cancer cell lines. , 2002, Journal of agricultural and food chemistry.

[3]  J. Siewert,et al.  Individualizing therapy in gastric cancer , 2003, Expert review of anticancer therapy.

[4]  S. Korsmeyer,et al.  Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death , 1993, Cell.

[5]  G. Capellá,et al.  Genetic susceptibility and gastric cancer risk , 2002, International journal of cancer.

[6]  J. Chung,et al.  Serotonin induces apoptosis in PGT-beta pineal gland tumor cells. , 2000, Japanese journal of pharmacology.

[7]  B. Patil,et al.  Differential Inhibition of Human Cancer Cell Proliferation by Citrus Limonoids , 2001, Nutrition and cancer.

[8]  V. Steele,et al.  Mechanistic considerations in the evaluation of chemopreventive data. , 1996, IARC scientific publications.

[9]  K. Grohmann,et al.  Biological properties of citrus flavonoids pertaining to cancer and inflammation. , 2001, Current medicinal chemistry.

[10]  K. Kim,et al.  Alcohol induces apoptosis in TM3 mouse Leydig cells via bax-dependent caspase-3 activation. , 2002, European journal of pharmacology.

[11]  Y. Tsujimoto,et al.  A cloning method for caspase substrates that uses the yeast two-hybrid system: cloning of the antiapoptotic gene gelsolin. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  J. Ferlay,et al.  Estimates of the worldwide mortality from eighteen major cancers in 1985. Implications for prevention and projections of future burden , 1993, International journal of cancer.

[13]  B. Rigas,et al.  Effect of aspirin on induction of apoptosis in HT-29 human colon adenocarcinoma cells. , 1998, Biochemical pharmacology.

[14]  S. Nagata,et al.  Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis , 1998, Nature.

[15]  A. Karande,et al.  Placental Protein 14 Induces Apoptosis in T Cells but Not in Monocytes* , 2001, The Journal of Biological Chemistry.

[16]  A. Spigelman,et al.  The variable effect on proliferation of a colon cancer cell line by the citrus fruit flavonoid Naringenin , 2003, Colorectal disease : the official journal of the Association of Coloproctology of Great Britain and Ireland.

[17]  M. Kobori,et al.  Flavonoids Inhibit Cell Growth and Induce Apoptosis in B16 Melanoma 4A5 Cells , 2000, Bioscience, biotechnology, and biochemistry.

[18]  Jeffrey S. Morris,et al.  Fish Oil Enhances Targeted Apoptosis During Colon Tumor Initiation in Part by Downregulating Bcl-2 , 2003, Nutrition and cancer.

[19]  N. Myong,et al.  The proteomics approach to find biomarkers in gastric cancer. , 2003, Journal of Korean medical science.

[20]  D. Forman,et al.  Gastric cancer, gastritis and plasma vitamin C: Results from an international correlation and cross‐sectional study , 1997, International journal of cancer.

[21]  A. Taraphdar,et al.  NATURAL PRODUCTS AS INDUCERS OF APOPTOSIS: IMPLICATION FOR CANCER THERAPY AND PREVENTION , 2001 .

[22]  P. Sylvester,et al.  Role of Caspase-8 Activation in Mediating Vitamin E-Induced Apoptosis in Murine Mammary Cancer Cells , 2003, Nutrition and cancer.

[23]  G M Cohen,et al.  Caspases: the executioners of apoptosis. , 1997, The Biochemical journal.

[24]  Patrick R. Griffin,et al.  Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis , 1995, Nature.

[25]  C. Napoli,et al.  Glycoxidation of Low‐Density Lipoprotein Increases TUNEL Positivity and CPP32 Activation in Human Coronary Cells , 2003, Annals of the New York Academy of Sciences.