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Correlation between ice activity and apoptosis induced by cisplatin in gastric cancer cell lines
A1402 AGA ABSTRACTS
GASTROENTEROLOGY Vol. 118, No.4
6368
6370
IN VITRO STUDIES INDICATE THAT NITROSATION IN ACID STOMACH WILL BE MAXIMAL AT GASTROESOPHAGEAL (GO) JUNCTION. Akihiko Moriya, Jeanette Grant, Andy Carswell, Craig Williams, Craig Mowat, Tom Preston, Sharon E. Anderson, Kenneth E. McColl, Univ of Glasgow, Glasgow, United Kingdom; Scottish Univ Research & Reactor Ctr, East Kilbride, United Kingdom. BACKGROUND: Nitrate absorbed from the GIT is secreted in saliva and converted to nitrite by buccal bacteria. Saliva is the main source of nitrite entering the acid stomach. Saliva also contains thiocyanate, a catalyst of nitrosation reactions. Ascorbic acid (AA) secreted in acidic gastric juice prevents the nitrite reacting with secondary amines to form nitrosoamines by converting it to nitric oxide (NO); in the process AA is oxidized to dehydroascorbic acid (DHA). AIMS AND METHODS: To study nitrosation under conditions simulating salivary delivery of nitrite into acidic gastric juice following a meal containing 2mmol nitrate (equivalent to salad portion). RESULTS: The delivery of nitrite (at a rate simulating salivary delivery) into HCL containing thiocyanate (Immol/I) resulted in nitrosation of morpholine to nitrosomorpholine. The rate of nitrosation was greatest at pH 2.5. Infusion of AA at a rate simulating its gastric secretion failed to prevent nitrosation. The ineffectiveness of the AA was due to oxygen in atmospheric air increasing by tenfold the amount of AA required to remove the nitrite by converting it all to NO. This increased requirement of AA was due to the NO combining with the oxygen to reform nitrite. Further studies showed that the rate of consumption of AA by acidified nitrite containing thiocyanate was extremely rapid in the presence of air with the nitrite consuming 5 times its molar equivalent of AA within 5 seconds. CONCLUSION: Air is swallowed along with nitrite and thiocyanate in saliva and will result in the rapid consumption of AA on entering the acid stomach and consequently uninhibited nitrosoamine formation. This will be maximal at the GO junction where the reactants first meet and may explain the high incidence of mutagenesis at this anatomical site.
CORRELATION BETWEEN ICE ACTIVITY AND APOPTOSIS INDUCED BY CISPLATIN IN GASTRIC CANCER CELL LINES.
6369 PREVENTION OF RADIATION-INDUCED PROCTITIS WITH GM-CSF VS MESALAZINE ENEMAS. A PROSPECTIVE, DOUBLE·BLIND STUDY. Ioannis A. Mouzas, Frosso Lyraraki, Christos Leontidis, Maria Tzardi, Dimitrios Haldeopoulos, Charalambos Varveris, Gastroenterology Dept, Med Sch, Univ of Crete, Heraklion, Greece; Radiotherapy Dept, Med Sch, Heraklion, Greece; Gastroenterology Dept, Heraklion, Greece; Pathology Dept, Med Sch, Heraklion, Greece; Radiotherapy Dept, Heraklion, Greece. There are no reports on treatment or prevention of radiation proctitis with enemas of granulocyte-macrophage colony-stimulating factor (GM-CSF). AIM:. To determine whether a GM-CSF solution can prevent radiationinduced proctitis in comparison to a known mesalazine solution, administered both as an enema. PATIENTS AND METHODS: Twenty-one patients beginning a cycle of pelvic radiation therapy (3000-5200 cGy), entered the present study.Following stratification, they were randomized, in a double-blind manner, to receive either a mesalazine (4 g/60 ml, once daily, 9 pat.) or an identical-appearing GM-CSF (400 mcg/60 ml, once daily, 12 pat.) solution during the 3-weeks period of pelvic radiation. In all patients a sigmoidoscopy was performed with biopsies from rectum and sigmoid before and following treatment. Sigmoidoscopy and histology were both classified according to a score ranging from 0 = normal to 3 = severe findings of proctitis. Clinical findings (bowel movements, consistence, rectal bleeding, tenesmus, feeling of incomplete evacuation), were assessed weekly before and during the course of the trial. A semiquantitative score was used ranging from 0 = normal findings to 3 = severe symptoms. RESULTS: Endoscopic, histological and clinical scores for both treatments are shown in the table. There were no significant differences between treatments. CONCLUSIONS: I. Topical GS-CSF enemas are safe and efficient for preventing radiation proctitis. 2. Topical GS-CSF and mesalazine are equally effective in reducing symptoms in radiationinduced proctitis.
Endoscopy, histology and clinical scores before and after treatment (GM-CSF vsMesalazine)
Endoscopy Histology GM·CSF/5· GM-CSG/5· ASA ASA before 1stweek 2nd week 3rdweek after treat
0210,3
0,510
11/13
1311
bowel mov
Clinical findings Conslsr. Tenestence bleeding mus
DID DID DID 05105 05 I 0,5
010 03106 03106 0,611 1.2/1.4
010 010 0/0 0103 DID
incompl evac
DID DID 09103 08113 0,9103 08113 0,9103 1.512 1.6113 1.612
Kazuya Muguruma, Bunzo Nakata, Masakazu Yashiro, Yoshito Yamashita, Nobuya Yamada, Yoshinari Ogawa, Naoyoshi Onoda, Tetsuji Sawada, Masaichi Ohira, Kosei Hirakawa. Osaka City Univ Med Sch, Osaka, Japan. Interleukin-I beta converting enzyme (ICE) has been reported to be related with anti-cancer drug induced apoptosis as well as CPP-32. To elucidate the ICE activity can be a predictor for the effect of chemotherapy, we examined the changes of ICE activity induced after exposure to cisplatin (CDDP) in gastric cancer cell lines. The high correlation between IC50 values and ICE activity ratio was shown (r=0.83, p=0.041).(ICE activity ratio: the ICE activity of cells at 4hr after CDDP treatment! the ICE Activity of no treated cells.) Furthermore, we examined the correlation between ICE activity and apoptosis induced by CDDP in the two cell lines which have very different CDDP sensitivities; OCUM-2M and OCUM2M1DDP, established in our labolatory from primary scirrhous gastric carcinoma. The apoptotic index of OCUM-2M was significantly higher than that of OCUM-2M (p=0.0005).On both Cell Lines, ICE activity started to increase immediately after exposure to CDDP and peaked at approximately 4hr after finishing exposure to CDDP and gradually decreased therefore. ICE activity of OCUM-2M was approximately 1.8-times higher than that of OCUM-2M1DDPat 4hr after exposure to CDDP. Taken together, our results indicate that to evaluate the changes of ICE activity after exposure to CDDP may be useful to predict apoptosis following CDDP treatment in gastric cancer cells.
6371 THE CORRELATION BETWEEN ANTICANCER EFFECT AND THE INHIBITORY EFFECT ON BONE-MARROW HEMOPOIE· SIS STEM CELL GROWTH AFTER DIFFERENTIAL CHEMO· THERAPY. Kazuya Muguruma, Bunzo Nakata, Masakazu Yashiro, Yoshito Yamashita, Nobuya Yamada, Naoyoshi Onoda, Tetsuji Sawada, Masaichi Ohira, Kosei Hirakawa, Osaka City Univ Med Sch, Osaka, Japan. (Introduction) FP therapy for the digestive cancer has been widely performed. We found the necessary cases to stop chemotherapy because of the bone marrow suppression. Recent studies have shown that the cell cycle of stem cells is rapid in comparison with that of the cancer cells. It has been also reported that it maintains the effect for the cancer and can relieve the side effect with the intermittent administration of the anticancer drug. In this study, we carried out the examination whether we can retain bone marrow activity, while the anticancer effect is maintained with the intermittent administration of the anticancer drug.(Material&Method) The side effect was examined by MTT assay method for bone-marrow hemopoiesis stem cell of infant peripheral blood. It was exposed at the 5-FU of double concentration of the continuous medication group for 4 days in the intermittent method to each well in adequate concentrarionin, area under curve was clinically equal, and the viable count was compared and was examined. And, it was similarly examined by the MTT assay method for cell line MKN-28 of the low-specialized gastric gland cancer derivation in the examination on the anticancer effect. To each well, 1000 cancer cells were injected, and were exposed at the concentration of 100 ng/ml and 200 ng/ml of 5-FU for 4 days, and the viable count was compared and was examined. (Result) The survival rate of hemopoietic stroma in continuously exposing at the concentration of 50 ng/ml of 5-FU was significantly lower than that of intermittent exposing of 5-FU (42.9:=7.3% vs 81.9:=13.2%), also in the concentration of 100 ng/ml and 500 ng/ml of 5-FU (p
GASTROENTEROLOGY Vol. 118, No.4
6368
6370
IN VITRO STUDIES INDICATE THAT NITROSATION IN ACID STOMACH WILL BE MAXIMAL AT GASTROESOPHAGEAL (GO) JUNCTION. Akihiko Moriya, Jeanette Grant, Andy Carswell, Craig Williams, Craig Mowat, Tom Preston, Sharon E. Anderson, Kenneth E. McColl, Univ of Glasgow, Glasgow, United Kingdom; Scottish Univ Research & Reactor Ctr, East Kilbride, United Kingdom. BACKGROUND: Nitrate absorbed from the GIT is secreted in saliva and converted to nitrite by buccal bacteria. Saliva is the main source of nitrite entering the acid stomach. Saliva also contains thiocyanate, a catalyst of nitrosation reactions. Ascorbic acid (AA) secreted in acidic gastric juice prevents the nitrite reacting with secondary amines to form nitrosoamines by converting it to nitric oxide (NO); in the process AA is oxidized to dehydroascorbic acid (DHA). AIMS AND METHODS: To study nitrosation under conditions simulating salivary delivery of nitrite into acidic gastric juice following a meal containing 2mmol nitrate (equivalent to salad portion). RESULTS: The delivery of nitrite (at a rate simulating salivary delivery) into HCL containing thiocyanate (Immol/I) resulted in nitrosation of morpholine to nitrosomorpholine. The rate of nitrosation was greatest at pH 2.5. Infusion of AA at a rate simulating its gastric secretion failed to prevent nitrosation. The ineffectiveness of the AA was due to oxygen in atmospheric air increasing by tenfold the amount of AA required to remove the nitrite by converting it all to NO. This increased requirement of AA was due to the NO combining with the oxygen to reform nitrite. Further studies showed that the rate of consumption of AA by acidified nitrite containing thiocyanate was extremely rapid in the presence of air with the nitrite consuming 5 times its molar equivalent of AA within 5 seconds. CONCLUSION: Air is swallowed along with nitrite and thiocyanate in saliva and will result in the rapid consumption of AA on entering the acid stomach and consequently uninhibited nitrosoamine formation. This will be maximal at the GO junction where the reactants first meet and may explain the high incidence of mutagenesis at this anatomical site.
CORRELATION BETWEEN ICE ACTIVITY AND APOPTOSIS INDUCED BY CISPLATIN IN GASTRIC CANCER CELL LINES.
6369 PREVENTION OF RADIATION-INDUCED PROCTITIS WITH GM-CSF VS MESALAZINE ENEMAS. A PROSPECTIVE, DOUBLE·BLIND STUDY. Ioannis A. Mouzas, Frosso Lyraraki, Christos Leontidis, Maria Tzardi, Dimitrios Haldeopoulos, Charalambos Varveris, Gastroenterology Dept, Med Sch, Univ of Crete, Heraklion, Greece; Radiotherapy Dept, Med Sch, Heraklion, Greece; Gastroenterology Dept, Heraklion, Greece; Pathology Dept, Med Sch, Heraklion, Greece; Radiotherapy Dept, Heraklion, Greece. There are no reports on treatment or prevention of radiation proctitis with enemas of granulocyte-macrophage colony-stimulating factor (GM-CSF). AIM:. To determine whether a GM-CSF solution can prevent radiationinduced proctitis in comparison to a known mesalazine solution, administered both as an enema. PATIENTS AND METHODS: Twenty-one patients beginning a cycle of pelvic radiation therapy (3000-5200 cGy), entered the present study.Following stratification, they were randomized, in a double-blind manner, to receive either a mesalazine (4 g/60 ml, once daily, 9 pat.) or an identical-appearing GM-CSF (400 mcg/60 ml, once daily, 12 pat.) solution during the 3-weeks period of pelvic radiation. In all patients a sigmoidoscopy was performed with biopsies from rectum and sigmoid before and following treatment. Sigmoidoscopy and histology were both classified according to a score ranging from 0 = normal to 3 = severe findings of proctitis. Clinical findings (bowel movements, consistence, rectal bleeding, tenesmus, feeling of incomplete evacuation), were assessed weekly before and during the course of the trial. A semiquantitative score was used ranging from 0 = normal findings to 3 = severe symptoms. RESULTS: Endoscopic, histological and clinical scores for both treatments are shown in the table. There were no significant differences between treatments. CONCLUSIONS: I. Topical GS-CSF enemas are safe and efficient for preventing radiation proctitis. 2. Topical GS-CSF and mesalazine are equally effective in reducing symptoms in radiationinduced proctitis.
Endoscopy, histology and clinical scores before and after treatment (GM-CSF vsMesalazine)
Endoscopy Histology GM·CSF/5· GM-CSG/5· ASA ASA before 1stweek 2nd week 3rdweek after treat
0210,3
0,510
11/13
1311
bowel mov
Clinical findings Conslsr. Tenestence bleeding mus
DID DID DID 05105 05 I 0,5
010 03106 03106 0,611 1.2/1.4
010 010 0/0 0103 DID
incompl evac
DID DID 09103 08113 0,9103 08113 0,9103 1.512 1.6113 1.612
Kazuya Muguruma, Bunzo Nakata, Masakazu Yashiro, Yoshito Yamashita, Nobuya Yamada, Yoshinari Ogawa, Naoyoshi Onoda, Tetsuji Sawada, Masaichi Ohira, Kosei Hirakawa. Osaka City Univ Med Sch, Osaka, Japan. Interleukin-I beta converting enzyme (ICE) has been reported to be related with anti-cancer drug induced apoptosis as well as CPP-32. To elucidate the ICE activity can be a predictor for the effect of chemotherapy, we examined the changes of ICE activity induced after exposure to cisplatin (CDDP) in gastric cancer cell lines. The high correlation between IC50 values and ICE activity ratio was shown (r=0.83, p=0.041).(ICE activity ratio: the ICE activity of cells at 4hr after CDDP treatment! the ICE Activity of no treated cells.) Furthermore, we examined the correlation between ICE activity and apoptosis induced by CDDP in the two cell lines which have very different CDDP sensitivities; OCUM-2M and OCUM2M1DDP, established in our labolatory from primary scirrhous gastric carcinoma. The apoptotic index of OCUM-2M was significantly higher than that of OCUM-2M (p=0.0005).On both Cell Lines, ICE activity started to increase immediately after exposure to CDDP and peaked at approximately 4hr after finishing exposure to CDDP and gradually decreased therefore. ICE activity of OCUM-2M was approximately 1.8-times higher than that of OCUM-2M1DDPat 4hr after exposure to CDDP. Taken together, our results indicate that to evaluate the changes of ICE activity after exposure to CDDP may be useful to predict apoptosis following CDDP treatment in gastric cancer cells.
6371 THE CORRELATION BETWEEN ANTICANCER EFFECT AND THE INHIBITORY EFFECT ON BONE-MARROW HEMOPOIE· SIS STEM CELL GROWTH AFTER DIFFERENTIAL CHEMO· THERAPY. Kazuya Muguruma, Bunzo Nakata, Masakazu Yashiro, Yoshito Yamashita, Nobuya Yamada, Naoyoshi Onoda, Tetsuji Sawada, Masaichi Ohira, Kosei Hirakawa, Osaka City Univ Med Sch, Osaka, Japan. (Introduction) FP therapy for the digestive cancer has been widely performed. We found the necessary cases to stop chemotherapy because of the bone marrow suppression. Recent studies have shown that the cell cycle of stem cells is rapid in comparison with that of the cancer cells. It has been also reported that it maintains the effect for the cancer and can relieve the side effect with the intermittent administration of the anticancer drug. In this study, we carried out the examination whether we can retain bone marrow activity, while the anticancer effect is maintained with the intermittent administration of the anticancer drug.(Material&Method) The side effect was examined by MTT assay method for bone-marrow hemopoiesis stem cell of infant peripheral blood. It was exposed at the 5-FU of double concentration of the continuous medication group for 4 days in the intermittent method to each well in adequate concentrarionin, area under curve was clinically equal, and the viable count was compared and was examined. And, it was similarly examined by the MTT assay method for cell line MKN-28 of the low-specialized gastric gland cancer derivation in the examination on the anticancer effect. To each well, 1000 cancer cells were injected, and were exposed at the concentration of 100 ng/ml and 200 ng/ml of 5-FU for 4 days, and the viable count was compared and was examined. (Result) The survival rate of hemopoietic stroma in continuously exposing at the concentration of 50 ng/ml of 5-FU was significantly lower than that of intermittent exposing of 5-FU (42.9:=7.3% vs 81.9:=13.2%), also in the concentration of 100 ng/ml and 500 ng/ml of 5-FU (p