食管癌引流淋巴结细胞诱导肿瘤细胞凋亡机制探讨

Mechanism of eophageal cancer cell apoptosis guided by human esophageal tumor-draining lymph node cell

目的探讨食管癌引流淋巴结(TDLN)细胞诱导肿瘤细胞凋亡的机制。方法将TDLN细胞分为3组进行培养。TDLN-Ⅰ组培养基中添加IL-2,TDLN-Ⅱ组添加IL-2+IL-4+GM-CSF,TDLN-Ⅲ组添加IL-2+IL-4+GM-CSF+自身食管癌细胞抗原(tAg)。用术中切取的食管癌组织块建立荷瘤裸鼠模型,随机分为空白对照组、IL-2组、TDLN-Ⅰ组、TDLN-Ⅱ组、TDLN-Ⅲ组,分别给予生理盐水、IL-2及相应TDLN细胞。利用流式细胞术检测瘤细胞凋亡率和Fas、FasL、Bax、Bc l-2、survivin蛋白的表达。结果对照组、IL-2组、TDLN-Ⅰ组、TDLN-Ⅱ组、TDLN-Ⅲ组肿瘤细胞的凋亡率分别为6.4%±3.1%、11.1%±1.3%、22.0%±2.8%、23.3%±1.9%、33.6%±5.2%,两两比较,P均<0.05;Fas蛋白的FI值分别为1.38±0.02、1.39±0.09、1.36±0.06、1.34±0.11、1.32±0.05,两组相比,P均>0.05;FasL蛋白的FI值分别为1.45±0.09、1.49±0.11、1.50±0.15、1.47±0.06、1.49±0.08,两组相比,P均>0.05;Bax蛋白的FI值分别为1.18±0.08、1.20±0.12、1.48±0.18、1.56±0.08、1.66±0.07。结论TDLN细胞可能通过影响Bax/Bc l-2和survivin蛋白表达诱导肿瘤细胞凋亡。

Objective To investigate the meehanism of the human esophageal tumor-draining lymph node（TDLN） cell guiding the tumor cell apoptosis. Methods The TDLN cells were cultured in three groups ： TDLN- Ⅰ group （ containing IL- 2） , TDLN- Ⅱ group （ containing IL-2, IL-4and GM-CSF） and TDLN- Ⅲ group （ containing IL-2, IL--4, GM-CSF and autologous tumor antigen）. A piece of fresh tumor specimen was obtained during operation and established tumor-baring nude mice model. The mice were randomized into 5 groups ： blank control group, IL-2 group, TDLN-Ⅰ group, TDLN- Ⅱ group and TDLN-Ⅲ group. The mice were injected NS, IL-2, TDLN cells respectively. The apoptotic ratio, Fas/Fasl, Bax/Bel- 2 and survivin in tumor tissue were detected by FCM. Results The apoptotie ratio of transplantation tumor in control group, IL-2 group, TDLN- Ⅰ group, TDLN- Ⅱ group and TDLN-Ⅲ group was 6.4% ±3.1%, 11.1% ± 1.3%, 22.0% ± 2.8%, 23.3% ±1.9 and 33.6% ±5.2%, respectively（P 〈0.05） ;Fas protein was 1.38 ±0.02, 1.39 ±0.09, 1.36± 0.06, 1.34 ±0.11 and 1.32 ±0. 05, respectively （P 〉0.05） ; FasL protein was 1.45 ±0.09, 1.49 ±0. 11, 1.50 ±0.15, 1.47±0.06 and l. 49 ± 0. 08 , respectively （P〉0.05）; Bax protein was 1.18±0.08, 1.20±0.12, 1.48 ± 0.18, 1.56 ± 0.08 and 1.66 ± 0.07, respectively. Conclusion TDLN ceils can induce apoptosis in transplanted tumor ceils by influence the expression of Bax/Bel-2 and survivin protein.