AIM： To explore the effect of gastrin 17 （G17） on β-catenin/T cell factor-4 （Tcf-4） signaling in colonic cancer cell line Colo320WT. METHODS： The pCR3.1/GR plasmid, which expresses gastrin receptor, cholecystok...AIM： To explore the effect of gastrin 17 （G17） on β-catenin/T cell factor-4 （Tcf-4） signaling in colonic cancer cell line Colo320WT. METHODS： The pCR3.1/GR plasmid, which expresses gastrin receptor, cholecystokinin-2 receptor （CCK-2R）, was transfected into a colonic cancer cell line Colo320 by Lipofectamine ^TM 2000 and the stably expressing CCK-2R clones were screened by G418. The expression levels of gastrin receptor in the Colo320 and the transfected Colo320WT cell line were assayed by RTPCR. Colo320WT cells were treated with G17 in a time-dependent manner （0, 1, 6, 12, 24 and 48 h）, then with L365,260 （Gastrin17 receptor blocker） for 30 rain, and with G17 again for 12 h or L365,260 for 12 h. Expression levels of β-catenin in a TX-100 soluble fraction and TX-100 insoluble fraction of Colo320WT cells treated with G17 were detected by co-immuniprecipation and Western blot. Immunocytochemistry was used to examine the distribution of β-catenin in CoLoWT320 cells. Expression levels of c-myc and cyclin D1 in Colo320WT cells treated with G17 were assayed by Western blot. RESULTS： Expression levels of β-catenin in the TX-100 solution fraction decreased apparently in a time- dependent fashion and reached the highest level after G17 treatment for 12 h, while expression levels of β-catenin in the TX-100 insoluble fraction were just on the contrary. Immunocytochemistry showed that β-catenin was translocated from the cell membranes into the cytoplasm and nucleus under G17 treatment. Expression levels of c-myc and cyclin D1 in the G17- treated Colo320WT cells were markedly higher compared to the untreated Colo320WT cells. In addition, the aforementioned G17-stimulated responses were blocked by L365,260.CONCLUSION： Gastrin17 activates β-catenin/Tcf-4 signaling in Colo320WT cells, thereby leading to over- expression of c-myc and cyclin D1.展开更多
AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confi...AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confirmed by real-time reverse transcription- polymerase chain reaction （PCR）. Western blotting and semi-quantitative immunohistochemistry were used to examine protein levels in human pancreatic cancer cell lines and pancreatic tissues, respectively, The role of αv integrin on gastrin-induced cell adhesion was examined using blocking anti-αv integrin monoclonal antibodies. Adherent cells were quantified by staining with crystal violet.RESULTS： Using a ＂cancer genes＂ array we identi- fied c^v integrin as a new gastrin target gene in human pancreatic cancer cells. A quantitative real-time PCR approach was used to confirm αv integrin gene expression. We also demonstrate that Src family kinases and the PI 3-kinase, two signalling pathways specifically activated by the CCK-2 receptor （CCK2R）, are involved in gastrin-mediated αv integrin expression. In contrast, inhibition of the ERK pathway was without any effect on αv integrin expression induced by gastrin. Our results also show that gastrin modulates cell adhesion via αv integrins. Indeed, in vitro adhesion assays performed on fibronectin show that gastrin significantly increases adhesion of pancreatic cancer cells. The use of blocking anti-αv integrin monoclonal antibodies completely reversed the increase in cell-substrate adhesion induced by gastrin. In addition, we showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, leads to the overexpression of αv integrin. This process may contribute to pancreatic turnout development observed in these transgenic animals.CONCLUSION： αv integrin is a new gastrin target in pancreatic cancer models and contributes to gastrin effects on cell adhesion.展开更多
目的:探讨胃蛋白酶原(pepsinogen,PG)、胃泌素-17(gastrin-17,G-17)、糖链抗原72-4(carbohydrate antigen 72-4,CA72-4)、癌胚抗原(carcinoembryonic antigen,CEA)联合检测在胃癌筛查中的应用价值。方法:选取2010-01~2015-12在我院收治...目的:探讨胃蛋白酶原(pepsinogen,PG)、胃泌素-17(gastrin-17,G-17)、糖链抗原72-4(carbohydrate antigen 72-4,CA72-4)、癌胚抗原(carcinoembryonic antigen,CEA)联合检测在胃癌筛查中的应用价值。方法:选取2010-01~2015-12在我院收治的经胃镜检查与组织病理学检查确诊的胃癌患者95例为胃癌组,胃良性病变患者100例为胃良性病变组,同时选取同期健康体检者100例为对照组。对3组的血清PG、G-17、CA72-4、CEA水平进行检测。结果:胃癌组的PGⅠ、PGⅠ/PGⅡ水平低于胃良性病变组与对照组,G-17、CA72-4、CEA水平高于胃良性病变组与对照组( P <0.05)。胃癌组血清PGⅠ、PGⅠ/PGⅡ、G-17、CA72-4、CEA以及5项联合检测阳性率均高于胃良性病变组与对照组( P <0.05, P <0.01)。5项联合检测灵敏度为87.37%,阴性预测值为93.88%,准确度为90.51%,均高于各指标单项检测( P < 0.05 )。结论: PGⅠ、PGⅠ/PGⅡ、G-17、CA72-4、CEA检测在胃癌的诊断中具有重要价值, 5项联合检测可显著提高胃癌诊断的灵敏度与准确度,可在胃癌筛查中推广使用。展开更多
基金Supported by the National Natural Science Foundation of China, No. 30470782
文摘AIM： To explore the effect of gastrin 17 （G17） on β-catenin/T cell factor-4 （Tcf-4） signaling in colonic cancer cell line Colo320WT. METHODS： The pCR3.1/GR plasmid, which expresses gastrin receptor, cholecystokinin-2 receptor （CCK-2R）, was transfected into a colonic cancer cell line Colo320 by Lipofectamine ^TM 2000 and the stably expressing CCK-2R clones were screened by G418. The expression levels of gastrin receptor in the Colo320 and the transfected Colo320WT cell line were assayed by RTPCR. Colo320WT cells were treated with G17 in a time-dependent manner （0, 1, 6, 12, 24 and 48 h）, then with L365,260 （Gastrin17 receptor blocker） for 30 rain, and with G17 again for 12 h or L365,260 for 12 h. Expression levels of β-catenin in a TX-100 soluble fraction and TX-100 insoluble fraction of Colo320WT cells treated with G17 were detected by co-immuniprecipation and Western blot. Immunocytochemistry was used to examine the distribution of β-catenin in CoLoWT320 cells. Expression levels of c-myc and cyclin D1 in Colo320WT cells treated with G17 were assayed by Western blot. RESULTS： Expression levels of β-catenin in the TX-100 solution fraction decreased apparently in a time- dependent fashion and reached the highest level after G17 treatment for 12 h, while expression levels of β-catenin in the TX-100 insoluble fraction were just on the contrary. Immunocytochemistry showed that β-catenin was translocated from the cell membranes into the cytoplasm and nucleus under G17 treatment. Expression levels of c-myc and cyclin D1 in the G17- treated Colo320WT cells were markedly higher compared to the untreated Colo320WT cells. In addition, the aforementioned G17-stimulated responses were blocked by L365,260.CONCLUSION： Gastrin17 activates β-catenin/Tcf-4 signaling in Colo320WT cells, thereby leading to over- expression of c-myc and cyclin D1.
文摘AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confirmed by real-time reverse transcription- polymerase chain reaction （PCR）. Western blotting and semi-quantitative immunohistochemistry were used to examine protein levels in human pancreatic cancer cell lines and pancreatic tissues, respectively, The role of αv integrin on gastrin-induced cell adhesion was examined using blocking anti-αv integrin monoclonal antibodies. Adherent cells were quantified by staining with crystal violet.RESULTS： Using a ＂cancer genes＂ array we identi- fied c^v integrin as a new gastrin target gene in human pancreatic cancer cells. A quantitative real-time PCR approach was used to confirm αv integrin gene expression. We also demonstrate that Src family kinases and the PI 3-kinase, two signalling pathways specifically activated by the CCK-2 receptor （CCK2R）, are involved in gastrin-mediated αv integrin expression. In contrast, inhibition of the ERK pathway was without any effect on αv integrin expression induced by gastrin. Our results also show that gastrin modulates cell adhesion via αv integrins. Indeed, in vitro adhesion assays performed on fibronectin show that gastrin significantly increases adhesion of pancreatic cancer cells. The use of blocking anti-αv integrin monoclonal antibodies completely reversed the increase in cell-substrate adhesion induced by gastrin. In addition, we showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, leads to the overexpression of αv integrin. This process may contribute to pancreatic turnout development observed in these transgenic animals.CONCLUSION： αv integrin is a new gastrin target in pancreatic cancer models and contributes to gastrin effects on cell adhesion.
文摘目的:探讨胃蛋白酶原(pepsinogen,PG)、胃泌素-17(gastrin-17,G-17)、糖链抗原72-4(carbohydrate antigen 72-4,CA72-4)、癌胚抗原(carcinoembryonic antigen,CEA)联合检测在胃癌筛查中的应用价值。方法:选取2010-01~2015-12在我院收治的经胃镜检查与组织病理学检查确诊的胃癌患者95例为胃癌组,胃良性病变患者100例为胃良性病变组,同时选取同期健康体检者100例为对照组。对3组的血清PG、G-17、CA72-4、CEA水平进行检测。结果:胃癌组的PGⅠ、PGⅠ/PGⅡ水平低于胃良性病变组与对照组,G-17、CA72-4、CEA水平高于胃良性病变组与对照组( P <0.05)。胃癌组血清PGⅠ、PGⅠ/PGⅡ、G-17、CA72-4、CEA以及5项联合检测阳性率均高于胃良性病变组与对照组( P <0.05, P <0.01)。5项联合检测灵敏度为87.37%,阴性预测值为93.88%,准确度为90.51%,均高于各指标单项检测( P < 0.05 )。结论: PGⅠ、PGⅠ/PGⅡ、G-17、CA72-4、CEA检测在胃癌的诊断中具有重要价值, 5项联合检测可显著提高胃癌诊断的灵敏度与准确度,可在胃癌筛查中推广使用。
