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目的研究聚集素抗LNCaP细胞凋亡的作用及聚集素反义寡核苷酸(AS-ODN)对肿瘤坏死因子-α(TNF-α)细胞毒性作用的影响。方法培养转染全长聚集素序列的LNCaP细胞(A)为实验组,野生型LNCaP细胞(L)及转染PIRES2-EGFP空载体的LNCaP细胞(M)为对照组,以20 ng/ml TNF-α进行诱导,MTT及ELISA法检测3组细胞增殖活性与凋亡程度；转染聚集素AS-ODN后A细胞分4组,①对照组：常规培养；②AS-ODN组：转染AS-ODN,培养液不含TNF-α；③TNF-α组：未转染AS-ODN,培养液含20ng/ml TNF-α；④TNF-α+AS-ODN组：转染AS-ODN,培养液含20ng/ml TNF-α,观察4组细胞增殖活性与凋亡程度的变化。结果L、M、A3组细胞增殖活性分别为0.84±0.03、0.85±0.04、0.95±0.03,L与M间差异无统计学意义(P＞0.05),L、M与A相比增殖活性显著降低(P值均＜0.01)。3组细胞ELISA吸光度值分别为0.59±0.04、0.62±0.03、0.33±0.04,L与M间差异无统计学意义(P＞0.05),但L、M与A相比凋亡程度显著增高(P值均＜0.01)。A细胞①～④组细胞增殖活性吸光度值分别为1.30±0.03,1.25±0.03,0.99±0.03,0.80±0.03,两两比较组间差异均有统计学意义(P值均＜0.05)；4组细胞凋亡程度吸光度值分别为0.02±0.00,0.21±0.02,0.63±0.07,1.16±0.04,两两比较组间差异均有统计学意义(P值均＜0.01)。结论转染并永久表达全长序列聚集素后,TNF-α对LNCaP细胞的细胞毒性作用显著降低,转染聚集素AS-ODN显著增强TNF-α的细胞毒性作用,聚集素具有抗LNCaP细胞凋亡的作用。
Objective To study the effect of agglutinin on the apoptosis of LNCaP cells and the effect of AS-ODN on the cytotoxicity of tumor necrosis factor-α (TNF-α). Methods LNCaP cells (A) transfected with full-length fusion sequences were cultured in vitro. LNCaP cells (L) and LNCaP cells (M) transfected with PIRES2-EGFP empty vector were used as control. -α, and the proliferation and apoptosis of the three groups were detected by MTT and ELISA; A cells were transfected with AS-ODN and divided into four groups: ①control group: conventional culture; ②AS-ODN group: AS- ODN in culture medium without TNF-α; ③TNF-α group: untransfected AS-ODN, culture medium containing 20ng / ml TNF-α; ④TNF-α + AS-ODN group: AS- 20ng / ml TNF-α, observe the changes of cell proliferation activity and apoptosis in the 4 groups. Results The proliferation activities of L, M and A3 groups were 0.84 ± 0.03,0.85 ± 0.04 and 0.95 ± 0.03, respectively. There was no significant difference between L and M groups (P> 0.05). The proliferation activity of L and M groups was significantly lower than that of A (P <0.01). The absorbance values of the three groups of cells were 0.59 ± 0.04, 0.62 ± 0.03 and 0.33 ± 0.04, respectively. There was no significant difference between L and M (P> 0.05), but the apoptosis of L and M was significantly higher than that of A Value <0.01). The cell proliferative absorbance of group A ~ (4) were 1.30 ± 0.03,1.25 ± 0.03,0.99 ± 0.03,0.80 ± 0.03 respectively, with significant difference between any two groups (all P <0.05). In group 4 The absorbance values of apoptosis were 0.02 ± 0.00, 0.21 ± 0.02, 0.63 ± 0.07 and 1.16 ± 0.04, respectively. There was significant difference between any two groups (P <0.01). CONCLUSION: The cytotoxic effect of TNF-α on LNCaP cells was significantly decreased after transfected and permanently expressing full-length sequence aggregates. Transfection with AS-ODN significantly increased the cytotoxicity of TNF-α. The role of death.