目的:分析细胞分裂周期CDCA基因家族成员在胃癌中的表达.方法:通过Oncomine等多个数据库分析CDCA基因家族在不同肿瘤组织中的表达,采用GEPIA平台进一步分析TCGA数据库中CDCA基因家族在胃癌组织中的表达.结果:在Oncomine数据库中共收集了2664个不同类型肿瘤的研究结果,CDCA基因家族成员表达差异有统计学意义的研究结果共有427个,其中,表达上调的392个,表达下调的35个;在胃癌中高表达的研究有38个,低表达的研究有0个.在GEPIA数据库中有正常胃组织211例,胃癌组织有408例,CDCA1、CDCA2、CDCA3、CDCA4、CDCA5、CDCA6、CDCA7和CDCA8基因在胃癌组织的表达水平均显著高于癌旁组织(P<0.05),其改变倍数分别为2.756、2.275、2.734、1.719、2.991、1.080、3.399、3.880倍.结论:CDCA基因家族在胃癌中高表达,可能与胃癌的发生发展密切相关,有望成为胃癌早期诊断的潜在标志物.

目的 研究精制保和颗粒(RBG)对溃疡性结肠炎(UC)的影响及其潜在机制.方法 采用随机数字表法将24只C57BL/6小鼠分为对照组、RBG组、DSS组和DSS+RBG组,每组各6只.采用饮用2％葡聚糖硫酸钠(DSS)的方法建立UC小鼠模型.RBG组自由饮用双蒸水的同时灌胃0.2 mL RBG(药物浓度为1.5 g/kg)11 d,每日1次;DSS组自由饮用2％DSS连续7 d后换回自由饮用双蒸水至11 d,同时灌胃等体积的双蒸水11 d,每日1次;DSS+RBG组自由饮用2％DSS连续7 d后换回自由饮用双蒸水至11 d,同时灌胃0.2 mL RBG 11 d,每日1次;对照组自由饮用双蒸水同时灌胃等体积的双蒸水11 d,每日1次.每日测量小鼠的体质量和便血情况,并每2天计算疾病活动指数(DAI)评分;干预后取结肠,测量结肠长度和体质量,并采用HE染色和电子光学显微镜观察结肠组织形态病理变化;免疫组化染色检测小鼠结肠组织中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达水平.结果 与对照组比较,DSS组小鼠的体质量明显减轻,DAI评分明显增加,结肠长度明显缩短,结肠体质量明显降低(P<0.05);而与DSS组比较,DSS+RBG组小鼠DAI评分明显降低,结肠长度明显变长,结肠体质量明显增加(P<0.05);与对照组比较,DSS组小鼠结肠组织中IL-6和TNF-α 水平显著升高(P均<0.05);与DSS组比较,DSS+RBG组小鼠结肠中IL-6和TNF-α 水平显著降低(P均<0.05).结论 抑制UC结肠组织中IL-6和TNF-α水平可能是RBG治疗UC的作用机制之一.

目的 观察精制保和颗粒(RBF)对人大肠癌细胞移植瘤生长的影响及对Bcl-2和Bax表达的调控作用.方法 通过对BALB/c裸鼠皮下种植人结肠癌HCT116细胞建立裸鼠荷瘤模型,根据瘤体的体积将裸鼠随机分为对照组和RBF组各5只,分别给予等体积的生理盐水和RBF溶液150 mg/(kg·d),连续灌胃12 d.通过游标卡尺和电子天平分别检测瘤体体积和重量,采用HE染色观察组织病理学改变,通过TUNEL染色检测细胞凋亡情况,采用免疫组化法检测移植瘤组织中PCNA、Bax和Bcl-2蛋白的表达水平.结果 与对照组比较,RBF组移植瘤瘤体的体积、重量均显著降低(P均<0.05);HE染色发现RBF组移植瘤组织出现坏死;免疫组化和TUNEL染色检测显示RBF组中PCNA表达明显降低和TUNEL阳性染色细胞数增多(P均<0.05);免疫组化检测显示RBF组移植瘤组织中抗凋亡蛋白Bcl-2的表达明显降低,而促凋亡蛋白Bax的表达明显升高(P均<0.05).结论 RBF干预抑制大肠癌移植瘤细胞的生长,通过调控Bcl-2和Bax表达促进细胞凋亡和抑制细胞增殖可能是其重要机制之一.

Objective:To explore the effect of betulinic acid (BA) on the growth and metastasis of colon cancer cell line HCT116 and the underlying possible mechanism.Methods:HCT116 cells were cultured with McCoy's 5A medium containing 10%fetal bovine serum and 1%penicillin/streptomycin mixed solution in a 37℃,5%CO2incubator. HCT116 cells were seeded at a density of 0.2×105cells/mL in a 96-well plate and cultured overnight. Then the cells were treated with different concentrations (0,10,20, and 40 μmol/L) of BA for 24,48 and 72 h, respectively. And then CCK8 was added to detect its absorbance (A value) at 450 nm. HCT116 cells were seeded at a density of 0.4×105cells/mL in a 12-well plate and cultured overnight. The cells were then collected by digestion and centrifugation, and stained with trypan blue after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h. The total number of cells was calculated by using a Countstar BioMed automatic cell counter. HCT116 cells were seeded at a density of 0.4×105cells/mL in a 12-well plate and cultured overnight. Then the cells were collected by digestion and centrifugation after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h.1.0×105cells/mL cell suspension was used for the Transwell experiment of cell migration and invasion, respectively. The effects of different concentrations of BA intervention for 48 h on the migration and invasion ability of HCT116 were analyzed. Western blot was used to detect the effect of BA on the protein expression of TGF-β and Smad2/3 in HCT116 cells after treated with BA for 48 h.Results:(1) CCK8 assay showed that the cell viability in each group was significantly reduced after treatment with different concentrations of BA for 24,48 and 72 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (P<0.05); The cell count results showed the number of cells in each group was remarkably decreased, and the differences were statistically significant compared with the 0 μmol/L group respectively (P<0.05). (2) Transwell assay showed that the number of cells that migrated or invaded into the membrane was profoundly inhibited in each group after treatment with different concentrations of BA for 48 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (P<0.05). (3) Western blot assay showed that the protein expression of TGF-β and Smad2/3 was obviously decreased in each group after treated with different concentrations of BA for 48 h compared with the 0 μmol/L group.Conclusion:BA can inhibit the growth and metastasis of colon cancer cells, and its mechanism may be achived by regulating the TGF-β/Smad signaling pathway.