裸鼠体内人腺样囊性癌细胞凋亡的研究

The study of apoptosis of salivary adenoid cystic carcinoma in nude mice

目的 在人涎腺腺样囊性癌 (salivaryadenoidcysticcarcinoma,SACC)裸鼠移植瘤模型上 ,观察重组人肿瘤坏死因子α(recombinedhumantumornecrosisfactor α ,rhTNF α)诱导凋亡的形态学特征及bax、bcl 2蛋白的表达情况。方法 将 6× 10 10 /L的SACC细胞悬液接种于裸鼠皮下建立动物模型后 ,按rhTNF α 10 0× 10 4IU /kg或 10× 10 4IU /kg瘤体内直接注射给药。采用光镜、电镜、流式细胞术及原位凋亡试剂检测盒观察凋亡的形态学变化 ;采用免疫组织化学观察bax、bcl 2的表达情况。结果 移植瘤细胞凋亡率明显高于对照组 ,差异有显著性 (P <0 .0 1)。凋亡细胞核消失后出现钙盐沉积 ,形成砂粒小体。凋亡细胞对bax、bcl 2蛋白的表达明显上升 (P <0 .0 5 )。结论 砂粒体钙化是TNF α诱导的SACC裸鼠移植瘤细胞凋亡的特征和归宿 ;TNF α诱导的细胞凋亡能够特异性增强bax、bcl

Objective To study the feature of apoptosis of salivary adenoid cystic carcinoma (SACC) induced by recombined human tumor necrosis factor-α(rhTNF-α) in nude mice, and to evaluate the related genes expression of apoptosis. Methods Twelve SPF grade 4～5 weeks old female Balb/c nude mice were selected in this study. SACC-83 cells were collected to 6×10 7 per milliliter and injected subcutaneously. Group A and B were experimental group which was given 100×10 4 IU/kg TNF-α or 10×10 4 IU/kg TNF-α respectively. Group C was only given normal saline and used as normal control. The investigations were adopted by using both light and transmission electron microscope (LM and TEM), flow cytometer and In Situ Cell Death Detection Kit. The evaluations of bax and bcl-2 expression were utilized by immunohistochemistry. Results The percentage of apoptosis of transplanted tumors was much higher than that of the control (P<0.01). Apoptotic cells were calcified and grit bodies were formed. Apoptotic cells expressed and contained significantly higher proportions of both bax and bcl-2 proteins (P<0.05). Conclusions It is suggested that calcification may be the obvious feature and the last outcome of the apoptosis of SACC transplanted tumors. Apoptosis induced by TNF-α can increase the expressions of bax and bcl-2.