不同压力CO2气腹对胃癌细胞增殖凋亡的影响

目的通过体外模拟气腹环境,观察CO2在不同压力下对胃癌细胞MKN-45增殖、凋亡的影响。方法以CO2为气体介质,在气腹机作用下维持密闭培养箱内压力为0、5、10、15mmHg,作用时间为4h。于处理后用血气分析仪检测培养液pH值;MTT法检测细胞增殖情况;通过流式细胞仪观察细胞凋亡指数变化;采用AnnexinV-FITC/PI双标染色、流式细胞仪测定凋亡细胞比例。结果处理结束后即刻检测,CO2组细胞培养液呈酸性,但在恢复正常培养后6h即恢复正常。MTT法检测细胞增殖变化,CO2组0、5、10mmHg压力下处理组与对照组比较无明显差异(P>0.05),而在15mmHg压力下处理组与对照组比较OD490值在前3d无明显差异(P>0.05),在4～7dOD490值下降非常显著(P<0.01)。在CO2环境下,0mmHg组细胞凋亡指数和凋亡比例与对照组相比无明显差异(P>0.05),而5、10、15mmHg组明显大于正常对照组(P<0.05)。结论在较低压力范围内(≤10mmHg)CO2对胃癌细胞增殖、凋亡影响不大,而在15mmHg压力下,CO2可抑制MKN-45细胞增殖,促进其凋亡。     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

目的 探讨不同压强持续性CO2气腹环境对胃癌细胞黏附与侵袭转移的影响.方法 建立体外CO2气腹模型,选用3种不同分化程度的胃癌细胞株MKN-45(低分化腺癌细胞)、SGC-7901(中分化腺癌细胞)和MKN-28(高分化腺痛细胞),分别在9 mm Hg、15 mm Hg以及常规条件下(0 mm Hg,对照组)作用2 h和4 h后,用RT-PCR法、CytoMatrixTM细胞黏附试剂盒和ECMatrixTM细胞侵袭试剂盒检测黏附分子E钙黏蛋白(E-cadherin)和细胞间黏附分子1(intercellular adhesionmolecule-1,ICAM-1)以及侵袭分子基质金属蛋白酶2(matrix metalloproteinases,MMP-2)和血管内皮生长因子A(vascular endothelial growth factor A,VEGF-A)的表达.将胃癌细胞注入裸鼠腹腔(2×106个细胞/只),每组10只.4周后每组取5只处死,记录腹腔成瘤情况,观察其余裸鼠的生存时间.结果 RT-PCR结果显示3种胃癌细胞株经CO2处理后,随着时程的延长及压强的升高,E-cadherin表达有下降的趋势(MKN-45:2.26→2.19、SGC-7901:2.16→2.09、MKN-28:2.06→1.99),而ICAM-1(MKN-45:2.20→2.28、SGC-7901:2.10→2.18、MKN-28:2.00→2.08)、MMP-2(MKN-45:2.05→2.13、SGC-7901:1.95→2.03、MKN-28:1.85→1.93)和VEGF-A(MKN-45:2.10→2.16、SGC-7901:2.00→2.06、MKN-28:1.90→1.96)则有升高的趋势,但是各实验组之间比较或实验组与对照组之间比较差异均无统计学意义(P>0.05).黏附侵袭实验也得出类似的结果.裸鼠模型显示3种胃癌细胞株在不同CO2气腹环境及对照条件下,腹腔成瘤个数随着时程的延长及压强的升高而增加(MKN-45:22→23、SGC-7901:20→22、MKN-28:21→22),存活天数则减少(MKN-45:23→21、SGC-7901:22→21、MKN-28:22→21),但各组的腹腔成瘤个数和存活时间之间相比差异均尤统计学意义(P>0.05).结论 在不高于15 mm Hg 压强以及不超过4 h的情况下,不同压强与时程的CO2对3种胃癌细胞株的黏附和侵袭能力并无显著影响,且不增加肿瘤的转移风险.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

CO2气腹环境对胃癌细胞黏附与侵袭转移的影响

Objective To investigate the influence of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expression of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cells MKN-45, SGC-7901 and MKN-28 were exposed to 3 different CO2 gradients: 9 mm Hg, 15 mm Hg and control group (0 mm Hg). The expression of E-cadherin, ICAM-1, MMP-2 and VEGF-A were measured at 2 and 4 hours exposure by using RT-PCR, CytoMatrixTM kit and ECMatrixTM kit. The pretreated gastric cancer cells were injected into abdominal cavity of nude mice(2×106 cells per mouse). Five mice in each group were sacrificed 4 weeks later to record the number of tumor nodules in abdominal cavity. The remaining mice were kept for observation of survival time. Results The expression of E-cadherin (MKN-45: from 2.26 to 2.19, SGC-7901 :from 2.16 to 2.09、MKN-28 :from 2.06 to 1.99), ICAM-1 (MKN-45 : from 2.20 to 2.28、SGC-7901: from 2.10 to 2.18、MKN-28: from 2.00 to 2.08), MMP-2 (MKN-45:from 2.05 to 2.13、SGC-7901: from 1.95 to 2.03、MKN-28: from 1.85 to 1.93) and VEGF-A(MKN-45 : from 2.10 to 2.16、SGC-7901 :from 2.00 to 2.06、MKN-28: from 1.90 to 1.96) didn't change significantly with increasing pressure and time (P>0.05). The expression of adhesive and invasive molecules didn't change significantly between the experimental groups and the control group. There was no statistical significance of tumor metastasis in abdominal cavity of nude mice(MKN-45:from 22 to 23、SGC-7901 :from 20 to 22、MKN-28:from 21 to 22) and survival time(MKN-45 :from 23 to 21、SGC-7901 :from 22 to 21、MKN-28 :from 22 to 21) among all the groups. Conclusion Under low pressure and short time of CO2 exposure, the adhesive and invasive capacity of gastric cancer cells did not change significantly hence did not increase the possibility of neoplasm metastasis.     

体外模拟二氧化碳气腹对人胃癌细胞迁移运动的影响

目的探讨通过体外模拟气腹环境，观察不同压力二氧化碳（CO2）和氦气（He）对胃癌细胞MKN-45迁移运动的影响。方法将MKN-45细胞置于充满CO2或He的密闭培养箱中，按模拟气腹种类不同分为对照组、CO2气腹组和He气腹组，模拟气腹压力分别12mm Hg和15mm Hg，作用时间均为4h。于处理后用血气分析仪检测培养液pH值；用Transwell法观察细胞迁移运动变化。结果处理结束后即刻检测，CO2组细胞培养液呈酸性，He组细胞培养液呈碱性。CO2组和He组在12mm Hg压力下MKN-45细胞穿过滤膜的数量较对照组差异无明显统计学（P〉0．05），CO2 15mm Hg组细胞穿过滤膜的数量较对照组明显减少（P〈0．01）；He 15mm Hg组与对照组差异无明显统计学（P〉0．05）。结论在临床常用气腹压力下（12mm Hg）CO2对胃癌细胞迁移运动无明显影响。     

不同压力的二氧化碳气腹对胃癌细胞迁移运动和细胞骨架的影响

目的通过体外模拟气腹环境，观察不同压力CO2或He（helium）对胃癌细胞MKN-45迁移运动和细胞骨架的影响。方法将MKN-45细胞置于充满CO2或He的密闭培养箱中，按模拟气腹种类不同分为对照组、CO2气腹组和He气腹组，模拟气腹压力分别为12mmHg和15mmHg，作用时间均为4h。于处理结束后即刻用血气分析仪检测培养液pH值；Transwell法观察细胞迁移运动变化；激光共聚焦显微镜观察细胞骨架变化。结果CO2组细胞培养液呈酸性，He组细胞培养液呈碱性。CO2组和He组在12mmHg压力下MKN-45细胞穿过滤膜的数量较对照组差异无统计学意义（P〉0．05）。CO2 15mmHg组细胞穿过滤膜的数量较对照组明显减少（P〈0．01）；He15mmHg组与对照组差异无统计学意义（P〉0．05）；CO2 15mmHg组细胞微丝、微管结构模糊，伪足消失。结论在临床常用气腹压力（12mmHg）下CO2对胃癌细胞迁移运动及细胞骨架无明显影响；在15mmHg压力下，CO2对胃癌细胞迁移运动起抑制作用，其作用机制可能与其细胞骨架受破坏有关。     

缺氧诱导因子1α对CO2气腹环境下人胃癌细胞凋亡的影响

目的 通过体外模拟CO2气腹环境,构建沉默缺氧诱导因子1α(HIF-1α)的RNAi表达载体,探讨HIF-1α对CO2气腹环境下人胃癌细胞MKN-45凋亡的影响及机制.方法 利用密闭培养箱模拟CO2气腹,气腹机维持培养箱内压力分别为0、5、10、15mm Hg.采用荧光定量RT-PCR方法和Western blot方法观察沉默HIF-1α前后MKN-45细胞HIF-1α mRNA和蛋白表达的变化;免疫细胞化学技术观察沉默HIF-1α前后细胞bcl-2/bax表达的变化;Annexin V-FITC/PI双标染色、流式细胞仪检测沉默HIF-1α前后细胞凋亡比例的变化.结果 沉默HIF-1α前15 mm Hg组细胞HIF-1αmRNA和蛋白表达(分别为1.51±0.04、4.44±0.30)均显著高于对照组(0.584±0.06、2.01±0.06)(P<0.01).沉默HIF-1α前15mmHg组细胞bcl-2/bax比值(0.77±0.05)较对照组(1.43±0.02)明显降低(P<0.05),细胞凋亡比例(11.60±2.11)较对照组(0.30±0.02)增加.而在沉默HI-1α后15 mm Hg组细胞HIF-1α mRNA(0.464±0.04)和蛋白表达(0.92±0.02)、bcl-2/bax比值(1.61±0.04)、细胞凋亡比例(0.4±0.03)与对照组相比差异均无统计学意义(P>0.05).结论 在CO2气腹环境压力为0、5、10mm Hg CO2时MKN-45细胞凋亡比例与对照组比较无差异,压力为15mm Hg时CO2气腹可促进胃癌细胞凋亡.HIF-1α可能为促进细胞凋亡的原因之一.     

CO2气腹对胃癌细胞周期及细胞周期蛋白的影响

目的 通过检测不同压力CO2气腹环境下胃癌细胞周期及其相关蛋白表达的变化,探讨CO2气腹对胃癌细胞生长增殖的影响.方法 将胃癌MKN-45细胞置于0、10、12和15 mm Hg CO2气腹环境下培养4 h,然后用流式细胞法检测胃癌细胞周期的变化,再用Western blot方法检测细胞周期蛋白Cyclin D1、CDK4、Rb和pRb的表达,最后用免疫沉淀法检测细胞Cyclin D1/CDK4结合力.结果 0、10、12 mm Hg CO2气腹组胃癌细胞增殖指数与对照组比较差异均无统计学意义(P>0.05),15 mm Hg CO2气腹组胃癌细胞增殖指数(27.4%4±3.7%)与对照组(36.4%±3.3%)比较显著下降(P<0.05).0、10、12 mm Hg CO2气腹组CDK4、Cyclin D1、pRb蛋白表达以及Cyclin D1和CDK4的结合能力与对照组比较差异均无统计学意义(P>0.05),15 mm Hg CO2气腹组CDK4、Cyclin D1、pRb蛋白表达以及Cyclin D1和CDK4的结合能力(0.71%±0.12%、0.93%±0.21%、0.54%±0.11%、0.18%±0.02%)与对照组(1.05%±0.16%、1.40%±0.24%、0.75%±0.14%、0.31%±0.02%)比较显著下降(P<0.05).0、10、12、15 mm Hg CO2气腹组Rb蛋白表达与对照组比较差异无统计学意义(P>0.05).结论 临床常用压力的CO2气腹对胃癌细胞周期无显著影响,15 mm Hg CO2气腹抑制细胞增殖,其原因可能与Cyclin D1和CDK4的表达下调有关.     

重组人内皮抑素联合应用5氟脲嘧啶对胃癌裸鼠移植瘤的影响

目的探讨重组人内皮抑素(recombinant human endostatin,rhES)与5氟脲嘧啶(5-fluorouracil,5-FU)联合应用对胃癌裸鼠移植瘤生长的抑制作用.方法建立胃癌异位移植BALB/C裸鼠模型,分为4组,每组6只.分别注射生理盐水,腹腔内注射5-FU(10 mg/kg),rhES组瘤周注射rhES(2 mg/kg)同时给予5-FU与rhES,每天1次,连用10 d.计算肿瘤体积、抑瘤率及肿瘤缩小率,检测肿瘤组织血管内皮生长因子(VEGF)、碱性成纤维生长因子(bFGF)、血管内皮生长因子-C(VEGF-C)、Ⅷ因子相关抗原(FⅧAg)、增殖细胞核抗原(PCNA)、bcl-2表达及肿瘤细胞凋亡指数(AI).结果rhES+5-FU组肿瘤体积为(43±2)mm3,5-FU组为(169±45)mm3,rhES组为(95±28)mm3,对照组为(1057±114)mm3(P＜0.01).抑瘤率为99.6%,肿瘤缩小率为98.2%.用药前rhES+5-FU组肿瘤体积为(207±50)mm3,比5-FU组与rhES组下降更迅速(P＜0.01).rhES+5-FU组与5-FU组VEGF、bFGF及VEGF-C表达强度均为0～+;rhES+5-FU组PCNA及bcl-2表达最弱;rhES+5-FU组AI为11.7±1.1,5-FU组为6.2±0.6,rhES组为5.8±0.8,对照组为2.4±0.6(P＜0.01).微血管密度在rhES+5-FU组为8.9±2.5,rhES组为10.0±1.5,均低于5-FU组(27.3±1.7)与对照组(29.9±2.3)(P＜0.01).结论联合应用5-FU及rhES能显著抑制胃癌血管生成,增加肿瘤细胞凋亡,使胃癌裸鼠移植瘤的肿瘤体积明显缩小.     

脱氧氟尿苷诱导胃腺癌细胞凋亡的临床研究

目的　了解术前化疗对胃腺癌细胞凋亡的影响。方法　采用抽签法术前随机分为口服脱氧氟尿苷(5′- DFUR)组(18例)、静脉输注5- FU CF组(16例)及对照组(2 0例)。化疗结束后第1～2天手术。采用免疫组织化学及TUNEL法检测癌细胞增殖指数(PI)及凋亡细胞指数(AI)。结果　PI在5′-DFUR组(40±13)、5 - FU CF组(41±15 )明显低于对照组(5 8±16 ) ,差异有统计学意义(P =0 . 0 0 0 ) ;AI在5′- DFUR组(14±9)、5 - FU CF组(14±10 )明显高于对照组(7±7) ,差异有统计学意义(P =0 . 0 17)。4 4例获得随访。总的0 . 5、1、2年生存率分别为93%、86 %、70 % ,0. 5、1、2年无瘤生存率分别为89%、77%、6 7%。3组术后生存时间差异无统计学意义。结论　术前3～5d短程口服5′- DFUR化疗可诱导胃腺癌细胞凋亡,降低癌细胞增殖指数,但不能改善术后生存情况。     

热CO2气腹对结肠癌细胞的增殖抑制作用及其机制

目的:探讨热CO2气腹对结肠癌细胞的增殖抑制作用及作用机制,明确其用于结直肠癌腹膜转移治疗的可行性。方法:建立热CO2气腹体外实验模型,热CO2气腹(43℃,2～4h)作用于结肠癌细胞株COLO205细胞。以WST-8法检测细胞增殖,Annexin-V/PI流式细胞术及透射电镜检测细胞死亡,流式细胞术检测细胞周期。结果:热CO2气腹(43℃,2～4h)对结肠癌细胞有显著的增殖抑制作用,热CO2显著诱导细胞凋亡及G1期细胞阻滞。结论:热CO2气腹通过诱导细胞凋亡及G1期细胞周期阻滞显著抑制结肠癌细胞的增殖,热CO2气腹具有应用于结直肠癌腹膜种植转移治疗的潜能。     

诱导型一氧化氮合酶抑制剂对胃癌MFC细胞增殖与凋亡的影响

目的研究诱导型一氧化氮合酶(iNOS)选择性抑制剂氨基胍(AG)对鼠胃癌MFC细胞增殖和凋亡的影响。方法将40只MFC胃癌皮下接种模型小鼠随机分成4组对照组(等体积生理盐水)、丝裂霉素组(MMC,每周注射2次,每次0.7mg/kg)、AG组(150mg·kg-1·d-1)和MMC加AG组。均采用腹腔内注射给药法,持续14d;停药后24h处死动物。应用Greiss反应法测定荷瘤动物血浆中一氧化氮(NO)量;剥离肿瘤,称重并计算抑瘤率;应用免疫组织化学法检测肿瘤细胞中增殖细胞核抗原(PCNA)的表达,TdT介导的dUTP缺口末端标记(TUNEL)法检测肿瘤细胞的凋亡,透射电镜观察凋亡细胞的超微结构。结果AG显著降低荷瘤小鼠血浆中NO的合成,明显抑制肿瘤的生长,抑瘤率为47.1%,与对照组比较,P<0.01。AG组的肿瘤细胞PCNA表达值为25.2±3.6,MMC加AG组为22.0±3.6,明显低于对照组的82.0±3.6(P<0.01);但凋亡指数未见下降,电镜下亦未见典型的细胞凋亡形态学变化。结论AG通过抑制肿瘤组织中NO的产生而抑制肿瘤生长,但并未促进肿瘤细胞凋亡。