肿瘤间质成纤维细胞对体外培养唾液腺多形性腺瘤细胞生物学行为的影响

doi:10.7518/hxkq.2023.2022314

华西口腔医学杂志 ›› 2023, Vol. 41 ›› Issue (2): 149-156.doi: 10.7518/hxkq.2023.2022314

肿瘤间质成纤维细胞对体外培养唾液腺多形性腺瘤细胞生物学行为的影响

侯亚丽¹(), 李荷香¹, 宋鹏¹, 杨艳霄¹, 郝亚丽², 刘慧娟²()

^1.河北医科大学口腔医（学）院病理科，河北省口腔医学重点实验室，河北省口腔疾病临床医学研究中心，石家庄 050017
^2.河北医科大学口腔医（学）院重点实验室，河北省口腔医学重点实验室，河北省口腔疾病临床医学研究中心，石家庄 050017

收稿日期:2022-08-16 修回日期:2023-01-22 出版日期:2023-04-01 发布日期:2023-04-14
通讯作者: 刘慧娟 E-mail:kqyyhyl@163.com;lhj2012001@163.com
作者简介:侯亚丽，副主任医师，硕士，E-mail：kqyyhyl@163.com
基金资助:
河北省医学科学研究课题(20210349);河北医科大学大学生创新性实验计划项目(USIP2021110)

Effect of tumor-stromal fibroblasts on the biological behavior of salivary gland pleomorphic adenoma cells in vitro

Hou Yali¹(), Li Hexiang¹, Song Peng¹, Yang Yanxiao¹, Hao Yali², Liu Huijuan²()

^1.Dept. of Pathology, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, China
^2.Key Laboratory of Stomatology, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, China

Received:2022-08-16 Revised:2023-01-22 Online:2023-04-01 Published:2023-04-14
Contact: Liu Huijuan E-mail:kqyyhyl@163.com;lhj2012001@163.com
Supported by:
Medical Science Research Project of Hebei Province(20210349);Hebei Medical University Undergraduate Innovation Experiment Project(USIP2021110);Correspondence: Liu Huijuan, E-mail: lhj2012001@163.com

摘要/Abstract

摘要：

目的探讨肿瘤间质成纤维细胞（TSF）对体外培养唾液腺多形性腺瘤（SPA）细胞增殖、侵袭和迁移的影响。方法采用组织块培养法培养唾液腺多形性腺瘤细胞（SPAC）、TSF和瘤旁正常成纤维细胞（NF），并通过免疫细胞化学染色法进行细胞鉴定。选取对数期TSF、NF制备条件培养液，培养SPAC并命名为TSF-SPAC组和NF-SPAC组，单纯培养SPAC作为对照组（SPAC组）。采用MTT实验、Transwell实验和划痕实验分别检测3组细胞的增殖、侵袭和迁移能力；酶联免疫吸附测定（ELISA）检测3组培养液中血管内皮生长因子（VEGF）的表达量。结果免疫细胞化学染色显示：波形蛋白（Vimentin）在TSF和NF中为阳性，而α-平滑肌肌动蛋白（α-SMA）和成纤维细胞活化蛋白（FAP）在NF中为阴性，在TSF中为弱阳性。MTT结果显示：TSF-SPAC组细胞增殖与NF-SPAC组和SPAC组的差异有统计学意义（P<0.05），NF-SPAC组与SPAC组的细胞增殖的差异无统计学意义（P>0.05）。Transwell侵袭实验和迁移实验显示：3组细胞的侵袭和迁移的差异无统计学意义（P>0.05）。ELISA检测结果显示：3组VEGF的表达量的差异均无统计学意义（P>0.05）。结论 TSF可能参与了SPA的生物学行为，促进体外培养的SPAC增殖，对其体外侵袭和迁移无促进作用，为SPA寻找新的治疗靶点提供了方向。

关键词: 唾液腺多形性腺瘤, 肿瘤间质成纤维细胞, 增殖, 侵袭, 迁移, 交界性肿瘤

Abstract:

Objective This study aims to investigate the effects of tumor-stromal fibroblasts (TSFs) on the proliferation, invasion, and migration of salivary gland pleomorphic adenoma (SPA) cells in vitro. Methods Salivary gland pleomorphic adenoma cells (SPACs), TSFs, and peri-tumorous normal fibroblasts (NFs) were obtained by tissue primary culture and identified by immunocytochemical staining. The conditioned medium was obtained from TSF and NF in logarithmic phase. SPACs were cultured by conditioned medium and treated by TSF (group TSF-SPAC) and NF (group NF-SPAC). SPACs were used as the control group. The proliferation, invasion, and migration of the three groups of cells were detected by MTT, transwell, and scratch assays, respectively. The expression of vascular endothelial growth factor (VEGF) in the three groups was tested by enzyme linked immunosorbent assay (ELISA). Results Immunocytochemical staining showed positive vimentin expression in NF and TSF. Results also indicated the weak positive expression of α-smooth muscle actin (SMA) and fibroblast activation protein (FAP) in TSFs and the negative expression of α-SMA and FAP in NFs. MTT assay showed that cell proliferation in the TSF-SPAC group was significantly different from that in the NF-SPAC and SPAC groups (P<0.05). Cell proliferation was not different between the NF-SPAC and SPAC groups (P>0.05). Transwell and scratch assays showed no difference in cell invasion and migration among the groups (P>0.05). ELISA showed that no significant difference in VEGF expression among the three groups (P>0.05). Conclusion TSFs may be involved in SPA biological behavior by promoting the proliferation of SPACs but has no effect on the invasion and migration of SPACs in vitro. Hence, TSF may be a new therapeutic target in SPA treatment.

Key words: salivary pleomorphic adenoma, tumor-stromal fibroblast, proliferation, invasion, migration, borderline tumor

中图分类号:

侯亚丽, 李荷香, 宋鹏, 杨艳霄, 郝亚丽, 刘慧娟. 肿瘤间质成纤维细胞对体外培养唾液腺多形性腺瘤细胞生物学行为的影响[J]. 华西口腔医学杂志, 2023, 41(2): 149-156.

Hou Yali, Li Hexiang, Song Peng, Yang Yanxiao, Hao Yali, Liu Huijuan. Effect of tumor-stromal fibroblasts on the biological behavior of salivary gland pleomorphic adenoma cells in vitro[J]. West China Journal of Stomatology, 2023, 41(2): 149-156.

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参考文献 15

1	Mhaidly R, Mechta-Grigoriou F. Role of cancer-associa-ted fibroblast subpopulations in immune infiltration, as a new means of treatment in cancer[J]. Immunol Rev, 2021, 302(1): 259-272.
2	Vennin C, Mélénec P, Rouet R, et al. CAF hierarchy driven by pancreatic cancer cell p53-status creates a pro-metastatic and chemoresistant environment via perlecan[J]. Nat Commun, 2019, 10(1): 3637.
3	Gao L, Morine Y, Yamada S, et al. The BAFF/NFκB axis is crucial to interactions between sorafenib-resistant HCC cells and cancer-associated fibroblasts[J]. Cancer Sci, 2021, 112(9): 3545-3554.
4	McAndrews KM, Chen Y, Darpolor JK, et al. Identification of functional heterogeneity of carcinoma-associated fibroblasts with distinct IL6-mediated therapy resistance in pancreatic cancer[J]. Cancer Discov, 2022, 12(6): 1580-1597.
5	Huang C, Liu J, He L, et al. The long noncoding RNA noncoding RNA activated by DNA damage (NORAD)-microRNA-496-Interleukin-33 axis affects carcinoma-associated fibroblasts-mediated gastric cancer development[J]. Bioengineered, 2021, 12(2): 11738-11755.
6	Espinosa CA, Fernández-Valle Á, Lequerica-Fernández P, et al. Clinicopathologic and surgical study of pleomorphic adenoma of the parotid gland: analysis of risk factors for recurrence and facial nerve dysfunction[J]. J Oral Maxillofac Surg, 2018, 76(2): 347-354.
7	Rooker SA, Van Abel KM, Yin LX, et al. Risk factors for subsequent recurrence after surgical treatment of recurrent pleomorphic adenoma of the parotid gland[J]. Head Neck, 2021, 43(4): 1088-1096.
8	Tang T, Huang X, Zhang G, et al. Advantages of targeting the tumor immune microenvironment over blocking immune checkpoint in cancer immunotherapy[J]. Signal Transduct Target Ther, 2021, 6(1): 72.
9	Mao X, Xu J, Wang W, et al. Crosstalk between cancer-associated fibroblasts and immune cells in the tumor microenvironment: new findings and future perspectives[J]. Mol Cancer, 2021, 20(1): 131.
10	Geng X, Chen H, Zhao L, et al. Cancer-associated fibroblast (CAF) geterogeneity and targeting therapy of CAFs in pancreatic cancer[J]. Front Cell Dev Biol, 2021, 9: 655152.
11	Zhou B, Chen WL, Wang YY, et al. A role for cancer-associated fibroblasts in inducing the epithelial-to-mesenchymal transition in human tongue squamous cell carcinoma[J]. J Oral Pathol Med, 2014, 43(8): 585-592.
12	王淑淑, 崔镓钰, 张凯佳, 等. SP13786通过CAFs外泌体抑制Stat3-EMT影响肺腺癌细胞A549的迁移侵袭 [J]. 中国肺癌杂志, 2021, 24(6): 384-393.
	Wang SS, Cui JY, Zhang KJ, et al. SP13786 inhibits the migration and invasion of lung adenocarcinoma cell A549 by supressing Stat3-EMT via CAFs exosomes[J]. Chin J Lung Cancer, 2021, 24(6): 384-393.
13	席磊, 付立新, 唐曦, 等. 癌相关成纤维细胞miR-200c 对乳腺癌细胞侵袭的影响[J]. 基因组学与应用生物学, 2018, 37(6): 2708-2713.
	Xi L, Fu LX, Tang X, et al. Effect of miR-200c in cancer-associated fibroblast on the invasion of human breast cancer cells[J]. Genom Appl Biol, 2018, 37(6): 2708-2713.
14	Spaeth EL, Dembinski JL, Sasser AK, et al. Mesenchymal stem cell transition to tumor-associated fibroblasts contributes to fibrovascular network expansion and tumor progression[J]. PLoS One, 2009, 4(4): e4992.
15	Melincovici CS, Boşca AB, Şuşman S, et al. Vascular endothelial growth factor (VEGF)-key factor in normal and pathological angiogenesis[J]. Rom J Morphol Embryol, 2018, 59(2): 455-467.

组别	细胞侵袭数	P值
NF-SPAC	21.30±5.10	0.612
TSF-SPAC	20.89±5.35
SPAC	23.20±5.51

组别	T0	T1	T0-T1	P值
NF-SPAC	1 770.17±17.42	1 570.17±27.58	200.00±34.00	0.098
TSF-SPAC	1 759.33±33.95	1 483.50±33.27	275.83±42.11
SPAC	2 107.67±27.27	1 774.00±33.03	333.67±49.65

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[10]	高永强, 施鹏伟, 师文楷, 刘一鸣. 长链非编码RNA HCG22在口腔鳞状细胞癌中的表达及作用机制研究[J]. 华西口腔医学杂志, 2021, 39(6): 658-666.
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肿瘤间质成纤维细胞对体外培养唾液腺多形性腺瘤细胞生物学行为的影响

Effect of tumor-stromal fibroblasts on the biological behavior of salivary gland pleomorphic adenoma cells in vitro

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