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目的如何诱导肿瘤细胞凋亡，是目前肿瘤治疗的研究热点之一．本研究旨在探讨肿瘤坏死因子α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒα，ＴＮＦα）诱导人大肠癌Ｌｏｖｏ细胞凋亡的可能性及其超微结构特点．为探索肿瘤治疗提供了新途径．方法Ｌｏｖｏ在体外用ＲＰＭＩ１６４０培养液、置５％ＣＯ２孵箱、３７℃常规培养．以ＴＮＦα终浓度１×１０４μ／ｍＬ作用２４，３６和４８ｈ，诱导Ｌｏｖｏ细胞凋亡．用倒置显微镜直接观察其光镜下形态改变；细胞经戊二醛和饿酸固定、丙酮脱水、环氧树脂包埋、超薄切片、醋酸铀－柠檬酸铅染色，用透射电镜观察其超微结构特点．结果①光镜下，随ＴＮＦα作用２４ｈ，Ｌｏｖｏ细胞由长梭形变为短梭状，胞体缩小；细胞间隙增宽，细胞密度无增加，生长抑制．作用３６ｈ，细胞变为椭圆形，少数短梭状；体积缩小，细胞内颗粒增多；仍贴壁生长．作用４８ｈ，细胞变为圆形，体积更小，颗粒较多，透亮度差；多数仍贴壁生长，少数上浮于液面．②电镜下，凋亡早期的Ｌｏｖｏ细胞外形不规则，胞体大小及胞质变化不大，微绒毛变细，染色质离散，开始向核膜下集中；中期，胞体缩小，胞膜完整，染色质呈颗粒、块状，凝集在核膜内侧、核孔只之间；后期：核裂解，形成凋亡小体?
Objective How to induce apoptosis of tumor cells is one of the research hotspots in cancer therapy. The purpose of this study was to investigate the possibility of apoptosis of human colon cancer Lovo cells induced by tumor necrosis factor α (TNFα) and its ultrastructural features. Provides a new way to explore cancer treatment. Methods Lovo was cultured in vitro with RPMI 1640 medium, 5% CO2 incubator, and 37°C routine culture. Apoptosis of Lovo cells was induced by 24, 36, and 48 h of TNFα at a final concentration of 1×104 μg/mL. The light microscope was used to observe the morphological changes directly; the cells were fixed with glutaraldehyde and hungry acid, dehydrated with acetone, embedded in epoxy resin, ultrathin section, uranyl acetate-lead citrate staining, and observed by transmission electron microscopy. Structural features. Results 1 Under light microscopy, Lovo cells changed from long spindle to short spindle with cell bodies shrinking with TNFα for 24 h; cell gaps were widened, cell density was not increased, and growth was inhibited. After 36 hours, the cells became elliptic and a few shuttlelike; the volume was reduced, intracellular particles increased; still adherent growth. After 48 hours, the cells became round, with smaller size, more particles, and poor brightness; most of them adhered to the wall and a few floated on the surface. 2 Under electron microscope, the appearance of Lovo cells with early apoptosis was irregular, cell body size and cytoplasm did not change, microvilli became fine, chromatin was discrete, and began to be concentrated under the nuclear membrane; in the middle stage, the cell body was shrinking and the cell membrane was intact. Particles, massive, agglutinated in the nuclear membrane, nuclear pores only; late: nuclear lysis, the formation of apoptotic bodies?