全反式维A酸可促进肿瘤相关诱导配体对多种胰腺癌细胞的凋亡作用

doi:10.16138/j.1673-6087.2023.03.007

摘要/Abstract

摘要：

目的: 观察全反式维A酸（all-trans retinoic acid，ATRA）是否能促进肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor related apoptosis induced ligand, TRAIL）对多种胰腺癌细胞的凋亡作用。方法: 将携带TRAIL基因的pCA13质粒分别转染SW1990、Patu8988和Bx-PC3等3种胰腺癌细胞，转染24 h后加入ATRA或等体积溶剂。四甲基偶氮唑盐（methyl thiazolyl tetrazolium，MTT）法测定细胞活性，流式细胞仪检测细胞的凋亡率和TRAIL受体R1、R2的表达，末端脱氧核苷酸转移酶（terminal deoxynucleotidyl transferase, TdT）介导的dUTP末端标记法（TdT mediated dUTP nick end labeling, TUNEL）和Hoechst双染色法、透射电镜观察细胞凋亡。结果: 转染pCA13/TRAIL质粒后再加入ATRA，细胞生长活性与未加ATRA组相比抑制显著（P<0.05）。流式细胞仪检测发现加入ATRA较未加药组明显促进细胞凋亡（P<0.05）。TUNEL和Hoechst双染色法后透射电镜可观察到细胞凋亡表现。但检测TRAIL-R1、TRAIL-R2的表达发现加入ATRA与未加药组差异无统计学意义（P>0.05）。结论: ATRA可促进TRAIL对多种胰腺癌细胞的凋亡作用，其机制与TRAIL-R1、TRAIL-R2的上调无关。

关键词: 全反式维A酸, 肿瘤坏死因子相关凋亡诱导配体, 胰腺癌, 凋亡

Abstract:

Objective To observe whether all trans retinoic acid (ATRA) can promote the apoptosis effect of tumor necrosis factor related apoptosis induced ligand (TRAIL) on various pancreatic cancer cells. Methods The pCA13 plasmid carrying TRAIL gene was transfected into three types of pancreatic cancer cell SW1990, Patu8988 and Bx-PC3 respectively. ATRA or equivalent solvent was added after 24 h of transfection. The cell viability was measured by the methyl thiazolyl tetrazolium(MTT) method, apoptosis rates and the expression of TRAIL receptor R1 and R2 were detected by flow cytometry, and the apoptosis was observed by terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick end labeling (TUNEL) and Hoechst double staining and transmission electron microscopy. Results The cell viability of the TRAIL+ATRA group was significantly inhibited compared to the group without ATRA (P<0.05). The result of the flow cytometry showed that ATRA significantly promoted cell apoptosis compared with the group without ATRA(P<0.05). Apoptosis was observed by transmission electron microscopy after TUNEL and Hoechst double staining. However, there was no significant difference in expression of TRAIL-R1 and R2 between the TRAIL+ATRA group and the non-ATRA group(P>0.05). Conclusions ATRA can promote the apoptosis of TRAIL on various pancreatic cancer cells, and its mechanism is not related to the up-regulation of TRAIL-R1 and R2.

Key words: All trans retinoic acid, Tumor necrosis factor related apoptosis induced ligand, Pancreatic cancer, Apoptosis

中图分类号:

R735.9

朱颖, 汤玉茗, 黄佳, 章永平, 姚玮艳. 全反式维A酸可促进肿瘤相关诱导配体对多种胰腺癌细胞的凋亡作用[J]. 内科理论与实践, 2023, 18(03): 171-176.

ZHU Ying, TANG Yuming, HUANG Jia, ZHANG Yongping, YAO Weiyan. All-trans retinoic acid promotes apoptosis effect of tumor necrosis factor related apoptosis induced ligand on a variety of pancreatic cancer cells[J]. Journal of Internal Medicine Concepts & Practice, 2023, 18(03): 171-176.

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参考文献 10

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细胞株转染时间	pCA13/TRAIL 质粒组	pCA13/TRAIL 质粒+ATRA组	t	P
SW1990
48 h	9.77±5.56	27.52±6.29	3.663 1	0.021 5
72 h	21.81±1.70	40.35±4.19	7.110 4	0.002 1
Patu8988
48 h	15.65±2.42	25.82±3.13	4.453 6	0.011 2
72 h	16.98±2.27	40.68±1.90	13.851 5	0.000 2
Bx-PC3
48 h	15.04±5.28	28.91±4.05	3.607 4	0.022 6
72 h	18.20±5.18	55.04±2.47	11.116 0	0.000 4

各细胞株作用时间	未转染组凋亡率	pCA13空质粒组凋亡率	pCA13/TRAIL质粒组凋亡率	空质粒+ATRA 组凋亡率	pCA13/TRAIL质粒+ATRA组凋亡率	F	P
SW1990
48 h	7.11±1.88	10.58±1.88^1）	27.30±5.14^1）	24.72±6.19	42.03±4.05	33.527	0.000 009
72 h	8.65±0.71	12.46±2.14^1）	31.45±4.93^1）	34.77±10.62	52.56±2.74	31.999	0.000 011
Patu8988
48 h	4.88±2.10	8.42±0.46^1）	13.52±0.95^1）	13.33±1.05	19.62±1.97	44.847	0.000 002
72 h	7.62±0.39	8.51±0.21^1）	14.43±3.13^1）	17.50±8.52	23.21±3.27	6.757	0.006 680
Bx-PC3
48 h	4.97±0.40	16.11±1.66^1）	31.40±8.70^1）	40.45±11.28	53.31±6.28	22.500	0.000 055
72 h	11.40±1.97	20.39±3.00^1）	43.48±5.76^1）	41.18±5.50	58.98±6.36	46.752	0.000 002

各细胞株作用时间	空质粒组	pCA13/TRAIL 质粒组	pCA13/TRAIL 质粒+ATRA组	P
SW1990
48 h	39.85±7.69	61.37±3.05	64.97±4.68	0.051
72 h	35.37±5.91	55.28±4.41	61.32±4.43	0.051
Patu8988
48 h	31.36±4.31	42.10±5.11	42.94±4.44	0.061
72 h	29.65±1.57	48.0±10.46	51.54±13.79	0.066
Bx-PC3
48 h	25.99±3.76	36.64±4.84	33.79±3.92	0.051
72 h	24.96±3.36	33.52±4.49	32.54±3.77	0.061

各细胞株作用时间	空质粒组	pCA13/TRAIL 质粒组	pCA13/TRAIL 质粒+ATRA组	P
SW1990
48 h	23.62±4.62	36.20±4.83^1）	39.03±3.24^1）	0.010
72 h	21.22±1.96	30.36±4.49^1）	38.03±6.14^1）	0.011
Patu8988
48 h	29.04±3.81	37.26±8.46	44.85±2.76^1）	0.037
72 h	28.01±3.36	42.32±9.25	49.66±8.01^1）	0.029
Bx-PC3
48 h	15.90±3.19	24.32±3.71^1）	28.88±3.11^1）	0.009
72 h	17.00±0.87	23.98±1.60^1）	27.33±4.73^1）	0.013

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