高压氧辅助健脾解毒方处理对体外培养人结肠癌SW620细胞恶性生物学行为的影响  

Effects of Jianpi Jiedu Formula assisted by hyperbaric oxygen on malignant biological behavior of SW620 human colon cancer cells cultured in vitro

作　　者：刘亮 韩秀庆 杜庆慧 张翔云 Liu Liang;Han Xiuqing;Du Qinghui;Zhang Xiangyun(Department of Traditional Chinese Medicine,Linyi Central Hospital,LinYi 276400,China)

机构地区：[1]临沂市中心医院中医科,临沂276400

出　　处：《中华航海医学与高气压医学杂志》2024年第1期101-106,共6页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

摘　　要：目的探讨高压氧(HBO)辅助健脾解毒方处理对体外培养人结肠癌SW620细胞恶性生物学行为的影响及其作用机制。方法体外培养人结肠癌SW620细胞完成后,按照实验设计分成4组:对照组、HBO组、健脾解毒方组和联合组。对照组SW620细胞不作如何处理,HBO组予以0.25 MPa HBO处理,健脾解毒方组予以1.50 mg/ml健脾解毒方制剂处理,联合组予以1.50 mg/ml健脾解毒方剂处理后再予以0.25 MPa HBO处理。首先在200倍倒置显微镜下观察SW620细胞形态的变化,采用MTT法检测SW620细胞的增殖能力,利用Hoechst 33258染色法检测SW620细胞凋亡情况,划痕实验检测SW620细胞迁移能力,Western blotting实验检测SW620细胞内凋亡蛋白的表达水平。结果处理48 h后,倒置显微镜下发现健脾解毒方组SW620细胞密度轻度减少,悬浮细胞稍微增多;联合组SW620细胞密度明显减少,部分细胞皱缩变圆,悬浮细胞明显增多(P<0.05)。Hoechst 33258荧光染色发现对照组和HBO组SW620细胞核形态和数量变化不明显;健脾解毒方组SW620细胞核呈致密浓染,形态异常,颜色发白,出现部分细胞凋亡;联合组SW620细胞核更加致密浓染,凋亡细胞明显增多(P<0.01);健脾解毒方组和联合组SW620细胞迁移能力受到明显抑制,联合组划痕愈合率明显高于健脾解毒方组(35.4%vs.48.9%,P<0.05);对照组、HBO组、健脾解毒方组SW620细胞G1期、S期和G2期变化不明显(P>0.05);而联合组SW620细胞G1期明显降低,S期明显增高(P<0.05),而G2期变化不明显(P>0.05);与对照组、健脾解毒方组、HBO组比较,联合组SW620细胞内凋亡蛋白Bax、caspase-3、Cytochrome C表达水平明显升高,Bcl-2蛋白表达明显降低(P<0.01)。结论HBO辅助健脾解毒方处理对SW620细胞具有抑制细胞增殖、诱导细胞凋亡及抑制细胞迁移的作用,其作用机制可能与下调Bcl-2蛋白表达,提高Bax、caspase-3、Cytochrome C蛋白表达水平有关。Objective To investigate the effect of Jianpi Jiedu Formula assisted by hyperbaric oxygen(HBO)on malignant biological behavior of SW620 human colon cancer cells cultured in vitro and its mechanism of action.Methods SW620 human colon cancer cells were cultured in vitro and divided into four groups according to the experimental design:control group,HBO group,Jianpi Jiedu Formula group,and combination group.The control group was not treated with any intervention,the HBO group was treated with 0.25 MPa HBO,the Jianpi Jiedu Formula group was treated with 1.50 mg/mL Jianpi Jiedu Formula preparation,and the combination group was treated with 1.50 mg/mL Jianpi Jiedu Formula preparation followed by 0.25 MPa HBO.The morphological changes of SW620 cells were observed under a 200×inverted microscope.MTT assay was used to access the proliferation ability of SW620 cells.Hoechst 33258 staining was conducted to detect the apoptosis of SW620 cells.Scratch test was used to evaluate the migration ability of SW620 cells.Western blotting was applied to measure the expression level of apoptin in SW620 cells.Results Forty-eight hours after treatment,in the Jianpi Jiedu Formula group,the density of SW620 cells was slightly decreased and the suspended cells were slightly increased under inverted microscope;in the combination group,the density of SW620 cells was significantly decreased,some cells were shrunk and rounded,and the number of suspended cells was significantly increased(P<0.05).Hoechst 33258 fluorescence staining showed no significant change in the morphology and the number of SW620 cell nuclei in the control group and the HBO group.The cell nuclei of SW620 cells in the Jianpi Jiedu Formula group were dense and densely stained,with abnormal morphology,white in color,and partial apoptosis,while those cell nuclei in the combination group were much denser and densely stained,with a significant increase in apoptotic cells(P<0.01).The cell migration abilities of SW620 cells in the Jianpi Jiedu Formula group and the combination grou

关 键 词：人结肠癌 SW620细胞 高压氧 健脾解毒方 体外培养 恶性生物学行为 

分 类 号：R735.35[医药卫生—肿瘤]