【作者】 李伟；

【导师】 王跃东；

【作者基本信息】 浙江大学 ， 外科学， 2002， 硕士

【摘要】 随着腹腔镜微创手术的不断发展，其应用范围日益扩大，现在包括腹腔镜辅助的结直肠恶性肿瘤切除术已逐渐在临床开展与应用。但一系列关于腹腔镜术后短期即出现戳孔处转移或腹膜播散的报道不断出现，使人们担心腹腔镜技术本身是否会促进癌细胞的生长和转移。因此，研究CO₂气腹对癌细胞本身的影响有一定的临床意义。本研究以模拟气腹处理结肠癌细胞后，通过处理后细胞生长周期变化及细胞表面粘附分子CD₄₄表达情况，来探讨CO₂气腹对体外培养的结肠癌细胞生长和转移潜能的影响。 材料和方法 采用由浙江大学肿瘤研究所提供的SW480结肠癌细胞株。以RPMI 1640培养液+10％小牛血清+青霉素100u/ml+链霉素100ug/ml作为培养液，常规放入37℃、5％CO₂恒温培养箱中培养。至一定数量后，分养入三块6孔板，每孔约2×10⁶个结肠癌细胞，随机分成CO₂组、Ar组和空白对照组，进行CO₂、Ar模拟气腹处理30分钟后，再行培养，检测处理后24h、48h、72h，结肠癌细胞的细胞周期及表面CD₄₄分子表达情况。 浙江大学硕十学位论文 结果与讨论 实验结果发现：CO。组 SW480结肠癌细胞在第 48h时，S期细胞数只有 7．2％，而 G；期细胞数达 89％，而在同期的空白组和 Ar组 S期细胞比率分别为37．2％、48．3％，G；期细胞比率为53．4％与47．5％。第72h时，CO。组S期细胞数有所回升达25厂％，但仍低于空白对照组和 Ar组3刁％与 64尸％人 G；期细胞数有所下降至 64．8％，仍高于空白对照组和 Ar组“1．20与 32．80人在CDM表达检测中，我们发现CO。组及Ar组经气腹处理后24h，表面CD。分子表达明显抑制，分别为 1．19％与 1石4％，明显低于对照 13．4％，但 48h、72h，CD44表达开始上升，Ar组分别为 15．2％与 20．9％，COZ组则更加明显分别达26．40与39．7o，超过了空白对照13．40。 以上结果表明：CO。气腹对体外培养的SW480结肠癌细胞有一定程度的抑制生长的作用，处理后第 48h时，S期细胞比率明显降低而G；期细胞比率明显升高，经统计学分析有显著性差异Pwto刀1。但这种抑制作用可能是短暂、可逆的，第 72h时CO。组的 S期比率即上升，G；期比率下降。而对CD刷分子表达来说，在第一个24h明显抑制，CD。。表达只有 1且9％，明显低 于对照 13．4％（Pwto刀1），而 48h。72h CD。。表达增高，达到 26．4％与 39．7％，高于对照水平，可能与转移有一定的关系。 结 论卜＊0。气腹对体外培养的结肠癌＊＊480细胞的生长有短暂可逆的 2 浙江人学硕1－学位论义 抑制作用。2、CO，气腹对体外培养的结肠癌SW480细胞的主长无促进作用3、CO。气腹对体外培养的结肠癌SW480细胞转移有一定的促进作用更多还原

【Abstract】 Numerous clinical reports have raised the possibility that laparoscopic cancer surgery, in contrast to conventional surgery, has an inherently detrimental effect on tumor growth. There have been numerous reports regarding site metastases and diffuse intraperitoneal metastases after laparoscopic surgery. So what is the influence of COi pneumoperitoneum on the cancer cells became a research point. The aim of this study was to examine the effects of CO2 pneumoperitoneum and Argon(Ar) pneumoperitoneum on the cytocyle and the CD44 expression in cultured SW480 colon cancer cells to explain potential dangers and risks of laparoscopic procedure.Materials and MethodsThe SW480 colon cancer cells were provided by the Zhejiang university tumor research center. We cultured the cell by RPMI 1640 solution in a culture box containing 5% COi at 37癈. Later, we divided the cells into three 6-well culture plates. Every well has 2X106 cells.Then we divided them into three groups randomly: CO2 group, Ar group and control group. We attempted to imitate pneumoperitoneum by increasing the ambient pressure to 15 mm mercury(Hg). The cells were exposed to the gases a total of 30 minutes. Then we measure the cyto cycle and CD44 expression at 24h, 48h and 72h after the effect of pneumoperitoneum.Result and DiscussionThe cell rate of S stage was obviously decreased in the CO2 group at 48h after the effect of pneumoperitoneum 7.2% compared with 48.3% in the Ar group , 37.2% in the control group. The percentage of stage Gl cells in the CO2 group was elevated to 89%,as compared to 47.5% of the Ar group and the 53.4% of control group(p<0.01). At 72h after the effect of pneumoperitoneum in the CO2 group, the cell rate of S stage was increased to 25.7% and the cell rate of Gl stage was decreased to 64.8%. In other words: CO2 pneumoperitoneum had no effect on promoting the growth of colon cancer cells, on the contrast it has some suppressive effect. As to the CD^ expression, it was obvious suppressed in the CO2 group and the Ar group 24h after the effect of pneumoperitoneum especially in the CO2 group 1.19% compared with 1.64% in the Ar group, 13.4% in the control group(p<0.01).But it was overexpressed in the CO2 group at the time of 48,72h after the effect of pneumoperitoneumwhich increased to 20.9% and 39.7% respectively as compared to 13.4% of the control group(p<0.01). From this experiment we can conclude that COi pneumoperitoneum has some effect on the colon cancer cell metastases ability. The mechanism need to be further studied.Conclusion1. CO2 pneumoperitoneum has no effect on promoting the growth of in vitro cultured colon cancer SW480 cells.2. CO2 pneumoperitoneum has positive effect on the metastases in vitro cultured colon cancer SW480 cells .更多还原