体外培养APA微囊化肿瘤坏死因子α/293细胞对结肠癌细胞增殖的影响

背景：积极探索结肠癌相关基因及抑癌基因已成为新的研究热点,人肿瘤坏死因子α是一个重要的促炎因子和免疫调节因子,对肿瘤的免疫治疗具有一定的疗效。目的：观察体外培养海藻酸钠-聚赖氨酸-海藻酸钠（alginate-polylysine-alginate,APA）微囊化肿瘤坏死因子α/293细胞对结肠癌细胞增殖的影响。方法：采用前期建立的制备方法,用APA微囊分别包裹人肿瘤坏死因子α/293细胞,APA微囊化0/293细胞。取对数生长期结肠癌（Lovo）细胞,配制成所需浓度的细胞悬液,接种24孔板培养,分别加入低、中、高剂量稳定转染的APA微囊化肿瘤坏死因子α/293,即分为5个实验组,APA微囊化肿瘤坏死因子α/293细胞低剂量组、中剂量组、高剂量组、阴性组为APA微囊化0/293细胞组,阳性组加入肿瘤坏死因子α,MTT法检测490nm吸光度;通过对人肿瘤细胞增殖抑制实验,观察对结肠癌细胞（Lovo）增殖的抑制作用。结果与结论：体外培养中,加入APA微囊化0/293细胞组对结肠癌细胞增殖无抑制作用;而APA微囊化肿瘤坏死因子α/293细胞中、高剂量组和阳性组,在24,48,72h的A值低于APA微囊化0/293细胞组（P〈0.05）,提示APA微囊化人肿瘤坏死因子α/293细胞所分泌肿瘤坏死因子α对结肠癌细胞有增殖抑制效应,均呈现出良好的数量依赖关系。

Tumor necrosis factor alpha-sercreting microcysts inhibit proliferation of Lovo colon cancer cells

BACKGROUND： This study was designed to establish gene engineered cells which auto-secreting human tumor necrosis factor-α （TNF-α）. Alginate-polylysine-alginate （APA） microencapsulated cells were co-cultured with colon cancer cells. On the one hand, the effects of micro-biological pump could be exerted to continously secrete TNF-α in the intracapsule cells. On the other hand, the immune rejection-prevented effects of APA microcapsules could be exerted to limit the overgrowth of highly-proliferative intracapsule cells. OBJECTIVE： To explore the effects of co-culture of TNFα-sercreting microcysts and human colon cancer cells Lovo on the proliferation of cancer cells. METHODS： Former established method was used. APA microcapsules were used to wrap TNF-α/293 cells and APA microencapsulated 0/293 cells. Logarithmic growth phase colon （Lovo） cells with DMEM containing 10% fetal bovine serum were prepared into cell suspension, and inoculated on 24-well plate. After 24 hours, the supernatant was discarded, and each well was added stable transfected APA microencapsulated TNF-α/293, which were divided into 5 groups including TNF-α/293 APA microencapsulated cell low dose group, middle dose group and high dose group, negative group and APA microencapsulated 0/293 cells. The positive group was added TNF-α factor, and MTT assay under the optical density 490 nm was applied. Through human tumor cell proliferation inhibition experiment, the inhibitory effects on colon cancer cells （Lovo） proliferation were observed. RESULTS AND CONCLUSION： APA microencapsulated 0/293 cell had no inhibitory effect on colon cancer cell proliferation in vitro, and there was no significant difference （P 0.05）. The middle, high-dose groups of APA microencapsulated TNF-α/293 cells and TNF-α-positive group showed a significant lower A at 24 h, 48 h, 72 h than the APA microencapsulated 0/293 cells group, with a significant difference （P 0.05）, suggesting that the middle, high-dose groups of APA microencapsulated TNF-α/293 cells and TNF-α-positive group had significant inhibition on the proliferation of colon cancer cells. The inhibition of APA microencapsulated TNF-α/293 cells secreted TNF-α on colon cancer cell proliferation shows a good dose-effect dependency, and expresses a considerable inhibition on the colon cancer cell proliferation as positive drug.