三乙醇胺对弥漫大B细胞淋巴瘤细胞生物特性的影响

张贵兵; 何正飞; 商文忠; 王爱伟; 吴艳芳; 黄惠燕

doi:10.16766/j.cnki.issn.1674-4152.002454

三乙醇胺对弥漫大B细胞淋巴瘤细胞生物特性的影响

doi: 10.16766/j.cnki.issn.1674-4152.002454

杭州市富阳区第一人民医院血液内科，浙江杭州 311400

基金项目:

浙江省中医药科技计划项目 2020ZA098

详细信息

通讯作者:
张贵兵，E-mail：qfp977@163.com

中图分类号: R733.1 R73-361
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- 文章访问数: 143
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- 被引次数: 0
出版历程
- 收稿日期: 2021-05-28
- 网络出版日期: 2022-09-05

Effect of triethanolamine on the biological characteristics of diffuse large B-cell lymphoma cells

Department of Hematdogy, the First People's Hospital of Fuyang District, Hangzhou City, Hangzhou, Zhejiang 311400, China

摘要

摘要: 目的观察三乙醇胺(TEOA)对弥漫大B细胞淋巴瘤(DLBCL)细胞生物特性的影响，旨在为DLBCL的临床治疗策略提供依据。方法 DLBCL细胞株OCI-LY3源于中国科学细胞库，随机数字表法简单随机分为对照组和观察组，每组分别设置5个复孔，观察组加入不同浓度(5、10、15、20、25、30、35、40、45、50 μmol/L)的TEOA于细胞培养箱培养12 h，采用Graph Pad Prism 5软件计算2组DLBCL细胞存活率，流式细胞术检测DLBCL细胞的凋亡和侵袭情况，MTT法检测DLBCL细胞的增殖情况，Western blotting检测蛋白相对表达量，比较2组细胞活性氧(ROS)水平, 收集2组研究数据并录入SPSS 20.0统计学软件进行分析。结果 TEOA作用后24、48、72 h观察组细胞增殖率为(39.24±3.95)%、(57.08±5.87)%、(114.08±14.69)%，明显低于对照组的(49.49±5.02)%、(95.12±9.64)%、(210.45±25.62)%，差异有统计学意义(均P＜0.05)；观察组穿膜细胞数[(38.14±3.77)个]明显低于对照组[(92.25±9.24)个]，而细胞凋亡率[(12.14±1.24)%]明显高于对照组[(3.64±0.36)%]，差异有统计学意义(P＜0.05)；TEOA可增加DLBCL OCI-LY3细胞内ROS的水平，并有效上调OCI-LY3中p38、p-Chk1、p-Chk2、γ-H2AX的表达水平。结论 TEOA可抑制细胞增殖和侵袭，同时有效诱导细胞凋亡及ROS的表达，TEOA作用于DLBCL OCI-LY3细胞可促进p38、p-Chk1、p-Chk2、γ-H2AX的表达，并诱导p38 MAPK通路的激活。
- 弥漫大B细胞淋巴瘤 /
- 三乙醇胺 /
- 细胞 /
- 生物特性
Abstract: Objective To observe the effect of triethanolamine (TEOA) on the biological characteristics of diffuse large B-cell lymphoma (DLBCL) cells, so as to provide a basis for clinical treatment of the disease. Methods DLBCL cell lines OCI-LY3 were provided by the cell bank of the Chinese Academy of Sciences. They were randomised into the control group and observation group by random number table method, with five parallel holes in each group. Different concentrations (5, 10, 15, 20, 25, 30, 35, 40, 45, 50 μmol/L) of TEOA were added into the observation group and then cultured for 12 h. The GraphPad Prism 5 software was used to calculate the survival rates of DLBCL cells in both groups, flow cytometry was used to detect the apoptosis and invasion of DLBCL cells, the MTT method was used to detect the proliferation of DLBCL cells, and Western Blotting was used to detect the relative protein expression. The levels of reactive oxygen species (ROS) in both groups were compared, and research data were collected and input into the SPSS 20.0 statistical software for statistical analysis. Results After 24, 48 and 72 h of TEOA treatment, the cell proliferation rates in the observation group[(39.24±3.95)%, (57.08±5.87)% and (114.08±14.69)%] were significantly lower than those in the control group[(49.49±5.02)%, (95.12±9.64)% and (210.45±25.62)%, all P < 0.05]. The observation group had significantly smaller number of transmembrane cells (38.14±3.77) and significantly higher apoptosis rate[(12.14±1.24)%] than the control group[92.25±9.24 and (3.64±0.36)%, respectively, all P < 0.05]. TEOA could increase ROS levels in DLBCL OCI-LY3 cells and effectively upregulate the expression levels of p38, p-Chk1, p-Chk2 and γ-H2AX in OCI-LY3. Conclusion TEOA can inhibit cell proliferation and invasion whilst effectively inducing cell apoptosis and ROS expression. TEOA acting on DLBCL OCI-LY3 cells can promote the expression of p38, p-Chk1, p-Chk2 and γ-H2AX and induce the activation of the p38 MAPK pathway.
- Diffuse large B-cell lymphoma /
- Triethanolamine /
- Cell /
- Biological characteristics

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图 1 TEOA作用后48 h DLBCL细胞增殖情况(HE染色，×200)

注：A为观察组；B为对照组。

Figure 1. Proliferation of DLBCL cells at 48 h after TEOA treatment (HE staining, ×200)

下载: 全尺寸图片幻灯片

表 1 不同时间TEOA对DLBCL细胞增殖的影响(x±s，%)

Table 1. Effects of TEOA on the proliferation of DLBCL cells at different time(x±s, %)

组别	n	24 h	48 h	72 h	F值	P值
观察组	5	39.24±3.95	57.08±5.87^a	114.08±14.69^a	226.491	＜0.001
对照组	5	49.49±5.02	95.12±9.64^a	210.45±25.62^a	529.343	＜0.001
t值		3.588	7.536	7.297
P值		0.007	＜0.001	＜0.001
注：与同组24 h比较，^aP＜0.05。

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表 2 TEOA对DLBCL细胞侵袭、凋亡的影响(x±s)

Table 2. Effects of TEOA on invasion and apoptosis of DLBCL cells(x±s)

组别	n	穿膜细胞数(个)	细胞凋亡率(%)
观察组	5	38.14±3.77	12.14±1.24
对照组	5	92.25±9.24	3.64±0.36
t值		12.124	14.720
P值		＜0.001	＜0.001

下载: 导出CSV

表 3 2组不同浓度TEOA作用后细胞内ROS水平变化(x±s)

Table 3. Changes of intracellular ROS levels in the two groups after treatment with different concentrations of TEOA(x±s)

组别	n	15 μmol/L	20 μmol/L	25 μmol/L	F值	P值
观察组	5	2.04±0.21	3.18±0.37^a	3.89±0.39^a	62.353	＜0.001
对照组	5	1.09±0.10	1.08±0.10	1.07±0.20	8.294	＜0.001
t值		9.133	12.252	14.387
P值		＜0.001	＜0.001	＜0.001
注：与同组15 μmol/L时的ROS水平比较，^aP＜0.05。

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表 4 TEOA诱导淋巴瘤细胞凋亡的分子机制分析(x±s，n=5)

Table 4. Molecular mechanism analysis of TEOA-induced apoptosis in lymphoma cells(x±s, n=5)

组别	时间	p38	Chk1	Chk2	γ-H2AX
观察组	干预前	0.32±0.02	0.21±0.02	0.27±0.03	0.30±0.03
	干预后	0.57±0.04	0.47±0.05	0.42±0.04	0.53±0.05
t值		-18.634	-16.611	-9.583	-12.857
P值		＜0.001	＜0.001	＜0.001	＜0.001

下载: 导出CSV

参考文献(20)

[1]	金静霞, 郑翠苹, 陈丽雅, 等. PD-1、PD-L1在弥漫大B细胞淋巴瘤组织中的差异性表达及其临床意义[J]. 临床血液学杂志, 2018, 31(1): 34-37. https://www.cnki.com.cn/Article/CJFDTOTAL-LCXZ201801011.htm JIN J X, ZHENG C P, CHEN L Y, et al. The differential expression levels and clinical significance of PD-1 and PD-L1 in tumor tissues of diffuse large B cell lymphoma[J]. Journal of Clinical Hematology, 2018, 31(1): 34-37. https://www.cnki.com.cn/Article/CJFDTOTAL-LCXZ201801011.htm
[2]	王友群, 唐小万. CHOP方案联合美罗华治疗弥漫大B细胞淋巴瘤78例回顾性分析[J]. 中华全科医学, 2018, 16(6): 916-918. doi: 10.16766/j.cnki.issn.1674-4152.000251 WANG Y Q, TANG X W. A retrospective analysis of 78 cases of diffuse large B-cell lymphoma treated with CHOP chemotherapy regimen combined with rituximab[J]. Chinese general practice, 2018, 16(6): 916-918. doi: 10.16766/j.cnki.issn.1674-4152.000251
[3]	CHEN T, YUAN Y, HUANG L S, et al. Dominant-negative PD1-armored CART cells induce remission in refractory diffuse large B-cell lymphoma (DLBCL) patients[J]. J Clin Oncol, 2019, 37(15): e19028.
[4]	KHURANA A, MWANGI R, NOWAKOWSKI G S, et al. Impact of organ function based standard exclusion criteria in diffuse large b-cell lymphoma (DLBCL) patients: Who gets left behind?[J]. J Clin Oncol, 2020, 38(15_suppl): e14100.
[5]	MONDELLO P, MIAN M. Frontline treatment of diffuse large B-cell lymphoma: Beyond R-CHOP[J]. Hematol Oncol, 2019, 37(4): 333-344.
[6]	王怡, 夏冰, 张翼鷟. 复发/难治性弥漫大B细胞淋巴瘤的分子诊疗进展[J]. 中国实验血液学杂志, 2018, 26(2): 603-608. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201802054.htm WANG Y, XIA B, ZHANG Y Z. Therapeutic progress in relapse/refractory diffuse large-B-cell lymphoma[J]. Journal of Experimental Hematology, 2018, 26(2): 603-608. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201802054.htm
[7]	李晓阳, 吴志平, 王梦馨, 等. 表没食子儿茶素没食子酸酯抗癌分子机制及其应用的研究进展[J]. 中草药, 2019, 50(13): 3217-3229. LI X Y, WU Z P, WANG M X, et al. Research progress on molecular mechanism of epigallocatechin-3-gallate against cancer and its application[J]. Chinese Traditional and Herbal Drugs, 2019, 50(13): 3217-3229.
[8]	张丹丹. TEOA抗人结肠癌细胞作用机制的初步研究[D]. 杭州: 浙江大学, 2017. ZHANG D D. Preliminary study on the mechanism of TEOA against human colon cancer cells[D]. Hangzhou: Zhejiang University, 2017.
[9]	DUBOIS S, JARDIN F. Novel molecular classifications of DLBCL[J]. Nat Rev Clin Oncol, 2018, 15(8): 474-476.
[10]	KAPADIA B, NANAJI N M, BHALLA K, et al. Fatty Acid Synthase induced S6Kinase facilitates USP11-eIF4B complex formation for sustained oncogenic translation in DLBCL[J]. Nat Commun, 2018, 9(1): 829.
[11]	林剑扬, 郑艳彬, 何鸿鸣, 等. DLBCL的临床病理特征及影响预后的相关因素分析[J]. 中国实验血液学杂志, 2018, 26(3): 779-783. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201803027.htm LIN J Y, ZHENG Y B, HE H M, et al. Clinicopathological features and prognostic factors of DLBCL[J]. Journal of Experimental Hematology, 2018, 26(3): 779-783. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201803027.htm
[12]	葛超, 吕梦迪, 张自由, 等. 基于天然产物的铂类和芳基金属抗癌药物研究进展[J]. 无机化学学报, 2020, 36(4): 597-606. https://www.cnki.com.cn/Article/CJFDTOTAL-WJHX202004002.htm GE C, LYU M D, ZHANG Z Y, et al. Trends of platinum and metal-arene anticancer drugs based on natural products[J]. Chinese Journal of Inorganic Chemistry, 2020, 36(4): 597-606. https://www.cnki.com.cn/Article/CJFDTOTAL-WJHX202004002.htm
[13]	TANG B, HUO Z, WU P. Study on a novel polyester composite nanofiltration membrane by interfacial polymerization of triethanolamine (TEOA) and trimesoyl chloride (TMC): I. Preparation, characterization and nanofiltration properties test of membrane[J]. J Membr Sci, 2008, 320: 198-205.
[14]	韩波, 高志棣, 王海霞, 等. miR-155在弥漫大B细胞淋巴瘤组织中的表达及其对细胞生物学特性的影响[J]. 中国实验血液学杂志, 2019, 27(2): 445-451. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201902026.htm HAN B, GAO Z L, WANG H X, et al. Expression of miR-155 in tissue of patients with diffuse large B-cell lymphoma and its effect on cell biological characteristics[J]. Journal of Experimental Hematology, 2019, 27(2): 445-451. https://www.cnki.com.cn/Article/CJFDTOTAL-XYSY201902026.htm
[15]	岳文君, 刘勇军, 陈京涛. NF-κB信号通路在弥漫大B细胞淋巴瘤中的作用及其靶向治疗中的应用[J]. 中国肿瘤生物治疗杂志, 2020, 27(1): 68-75. https://www.cnki.com.cn/Article/CJFDTOTAL-ZLSW202001012.htm YUE W J, LIU Y J, CHEN J T. The role of NF-κB signaling pathway in diffuse large B cell lymphoma and its application in targeted therapy[J]. Chinese Journal of Cancer Biotherapy, 2020, 27(1): 68-75. https://www.cnki.com.cn/Article/CJFDTOTAL-ZLSW202001012.htm
[16]	FU S, LUAN T, JIANG C Y, et al. miR-3622a promotes proliferation and invasion of bladder cancer cells by downregulating LASS2[J]. Gene, 2019, 701: 23-31.
[17]	刘靖, 杨画, 寻阳, 等. sCXCL16对弥漫大B细胞淋巴瘤的体外生物学影响与初步机制[J]. 临床与实验病理学杂志, 2019, 35(4): 393-397. https://www.cnki.com.cn/Article/CJFDTOTAL-LSBL201904004.htm LIU J, YANG H, XUN Y, et al. Effect of soluble CXCL16 on diffuse large B cell lymphoma in vitro and its preliminary mechanism[J]. Chinese Journal of Clinical and Experimental Pathology, 2019, 35(4): 393-397. https://www.cnki.com.cn/Article/CJFDTOTAL-LSBL201904004.htm
[18]	胡施炜, 楼恩哲, 王瑜, 等. 大黄酸通过抑制NF-κB通路促进DLBCL细胞OCI-LY8凋亡[J]. 中国病理生理杂志, 2019, 35(11): 1974-1980. https://www.cnki.com.cn/Article/CJFDTOTAL-ZBLS201911008.htm HU S W, LOU E Z, WANG Y, et al. Rhein promotes DLBCL cell OCI-LY8 apoptosis by inhibiting NF-κB signaling pathway[J]. Chinese Journal of Pathophysiology, 2019, 35(11): 1974-1980. https://www.cnki.com.cn/Article/CJFDTOTAL-ZBLS201911008.htm
[19]	HOJJAT-FARSANGI M, GHADERI A, DANESHMANESH A H, et al. Diffuse large B cell lymphoma (DLBCL) expresses ROR1 and a ROR1 small molecule inhibitor (KAN0441571C) induced significant apoptosis of tumor cells[J]. Blood, 2019, 134(Suppl 1): 2565.
[20]	余醒醒. TEOA通过激活ROS依赖的p38MAPK信号通路抑制弥漫性大B细胞淋巴瘤细胞的增殖并诱导其DNA损伤[D]. 蚌埠: 蚌埠医学院, 2019. YU X X. TEOA inhibits proliferation and induces DNA damage of diffuse large B-cell lymphoma cells by activating ROS-dependent p38MAPK signaling pathway[D]. Bengbu: Bengbu Medical College, 2019.