肿瘤相关巨噬细胞来源IL-6通过上调肿瘤细胞中LIF表达对口腔鳞状细胞癌Cal27细胞侵袭和迁移的促进作用

doi:10.13481/j.1671-587X.20220414

Abstract

Abstract: Background

To investigate the effects of tumor associated macrophages（TAMs）-derived interleukin-6 （IL-6） on the leukemia inhibitory factor （LIF） expression in oral squamous cell carcinoma （OSCC） Cal27 cells and the invasion and migration of OSCC Cal27 cells，and to clarify their possible mechanisms.

Methods

The human OSCC Cal27 cells and mouse peritoneal macrophages（Raw264.7 cells ）were cultured in vitro. The Raw264.7 cells were divided into control and experimental groups. The cells in control group were incubated with DMEM， and the cells in experimental group were incubated with Cal27 conditioned medium （Cal27-CM） to obtain the TAMs. The expression levels of IL-6 mRNA and IL-6 levels in the cells in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and enzyme-linked immunosorbent assay （ELISA） methods. The Cal27 cells were divided into control group and experimental group （5 μg·L^－1 Tocilizumab）.The Cal27 cells in control group were directly treated with TAMs conditioned medium， and the Cal27 cells in experimental group were treated with interleukin 6 receptor （IL-6R） inhibitor Tocilizumab in advance and then treated with TAMs conditioned medium. Transwell chamber experiment was used to detect the migration and invasion abilities of Cal27 cells in two groups. Cell scratch assay was used to detect the scratch healing rates of Cal27 cells in two groups. RT-qPCR and Western blotting methods were used to detect the expression of levels LIF mRNA and protein in Cal27 cells in two groups.

Results

The results of RT-qPCR and ELISA showed that the mRNA expression level of IL-6 and the IL-6 level in the TAMs in experimental group were significantly higher than those in the RAW264.7 cells in control group （P<0.01）. The results of Transwell chamber experiment showed that compared with control group， the number of migration and invasion cells in experimental group was significantly reduced （P<0.01）. The results of cell scratch experiment showed that compared with control group， the scratch healing rate of the Cal27 cells in experimental group was significantly decreased （P<0.01）. The RT-qPCR and Western blotting results showed that compared with control group， the expression levels of LIF mRNA and protein in the Cal27 cells in experimental group were decreased down-regulated （P<0.01）.

Conclusion

TAMs-derived IL-6 promotes the invasion and migration of OSCC Cal27 cells by up-regulating the expression of LIF in tumor cells.

Key words: Leukemia inhibitory factor, Oral squamous cell carcinoma, Tumor associated macrophages, Interleukin-6

CLC Number:

R739.8

Weibo LI,Juan CAO,Xiu GUO,Chunling DONG,Bo LI. Promotion effect of tumor associated macrophages-derived IL-6 on invasion and migration of oral squamous cell carcinoma Cal27 cells by upregulating LIF expression in tumor cells[J].Journal of Jilin University(Medicine Edition), 2022, 48(4): 946-953.

Figures/Tables 7

Tab.1

Fig.1

Fig.2

Tab.2

Fig. 3

Fig. 4

Tab.3

References 35

1	LU X L， CHEN L L， LI Y， et al. Long non-coding RNA LINC01207 promotes cell proliferation and migration but suppresses apoptosis and autophagy in oral squamous cell carcinoma by the microRNA-1301-3p/lactate dehydrogenase isoform A axis［J］. Bioengineered， 2021， 12（1）： 7780-7793.
2	YAO C， KONG F Z， ZHANG S L， et al. Long non-coding RNA BANCR promotes proliferation and migration in oral squamous cell carcinoma via MAPK signaling pathway［J］. J Oral Pathol Med， 2021， 50（3）： 308-315.
3	CHI A C， DAY T A， NEVILLE B W. Oral cavity and oropharyngeal squamous cell carcinoma： an update［J］. CA Cancer J Clin， 2015， 65（5）： 401-421.
4	SASAHIRA T， KIRITA T. Hallmarks of cancer-related newly prognostic factors of oral squamous cell carcinoma［J］. Int J Mol Sci， 2018， 19（8）： 2413.
5	MODESTO A， SIEGFRIED A， LUSQUE A， et al. Distinct outcomes of oropharyngeal squamous cell carcinoma patients after distant failure according to p16 status： implication in therapeutic options［J］. Curr Oncol， 2021， 28（3）： 1673-1680.
6	ZHONG Q， FANG Y X， LAI Q H， et al. CPEB3 inhibits epithelial-mesenchymal transition by disrupting the crosstalk between colorectal cancer cells and tumor-associated macrophages via IL-6R/STAT3 signaling［J］. J Exp Clin Cancer Res， 2020， 39（1）： 132.
7	LIAO Q， ZENG Z， GUO X， et al. LPLUNC1 suppresses IL-6-induced nasopharyngeal carcinoma cell proliferation via inhibiting the Stat3 activation［J］. Oncogene， 2014， 33（16）： 2098-2109.
8	SULLIVAN N J， SASSER A K， AXEL A E， et al. Interleukin-6 induces an epithelial-mesenchymal transition phenotype in human breast cancer cells［J］. Oncogene， 2009， 28（33）： 2940-2947.
9	SAITO K， MITSUI A， SUMARDIKA I W， et al. PLOD2-driven IL-6/STAT3 signaling promotes the invasion and metastasis of oral squamous cell carcinoma via activation of integrin β1［J］. Int J Oncol，2021，58（6）： 29.
10	WANG H Y， SI S N， JIANG M J， et al. Leukemia inhibitory factor is involved in the pathogenesis of NSCLC through activation of the STAT3 signaling pathway［J］. Oncol Lett， 2021， 22（3）： 663.
11	NICOLA N A， BABON J J. Leukemia inhibitory factor （LIF）［J］. Cytokine Growth Factor Rev， 2015， 26（5）： 533-544.
12	WYSOCZYNSKI M， MIEKUS K， JANKOWSKI K， et al. Leukemia inhibitory factor： a newly identified metastatic factor in rhabdomyosarcomas［J］. Cancer Res， 2007， 67（5）： 2131-2140.
13	LIU S C， TSANG N M， CHIANG W C， et al. Leukemia inhibitory factor promotes nasopharyngeal carcinoma progression and radioresistance［J］. J Clin Invest， 2013， 123（12）： 5269-5283.
14	YU H Y， YUE X T， ZHAO Y H， et al. LIF negatively regulates tumour-suppressor p53 through Stat3/ID1/MDM2 in colorectal cancers［J］. Nat Commun， 2014， 5： 5218.
15	LIN T A， WU T S， LI Y J， et al. Role and mechanism of LIF in oral squamous cell carcinoma progression［J］. J Clin Med， 2020， 9（2）： E295.
16	WANG W， TAN J Y. Co-inhibition of BET proteins and PD-L1 as a potential therapy for OSCC through synergistic inhibition of FOXM1 and PD-L1 expressions［J］.J Oral Pathol Med，2019，48（9）：817-825.
17	李蕴蕴，李瑞喆，秦慧，等. BAF53a表达对口腔鳞状细胞癌患者预后的影响［J］. 郑州大学学报（医学版）， 2020，55（4）： 512-515.
18	NAJAFI M， HASHEMI GORADEL N， FARHOOD B，et al. Macrophage polarity in cancer： a review［J］. J Cell Biochem， 2019， 120（3）： 2756-2765.
19	PETRUZZI M N， CHERUBINI K， SALUM F G，et al.Role of tumour-associated macrophages in oral squamous cells carcinoma progression： an update on current knowledge［J］. Diagn Pathol， 2017， 12（1）： 32.
20	HUANG C M， OU R Z， CHEN X N， et al. Tumor cell-derived SPON2 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by activating PYK₂ in CRC［J］. J Exp Clin Cancer Res， 2021， 40（1）： 304.
21	HUANG T T， SONG C L， ZHENG L， et al. The roles of extracellular vesicles in gastric cancer development， microenvironment， anti-cancer drug resistance， and therapy［J］.Mol Cancer，2019，18（1）： 62.
22	LIN Y X， XU J X， LAN H Y. Tumor-associated macrophages in tumor metastasis： biological roles and clinical therapeutic applications［J］. J Hematol Oncol， 2019， 12（1）： 76.
23	ZHAO X， QI X Q， LIAN W R， et al. Trichomicin suppresses colorectal cancer via comprehensive regulation of IL-6 and TNFα in tumor cells， TAMs， and CAFs［J］. Front Pharmacol， 2020， 11： 386.
24	GWAK J， JEONG H， LEE K， et al. SFMBT2-mediated infiltration of preadipocytes and TAMs in prostate cancer［J］.Cancers （Basel），2020，12（9）：E2718.
25	JIANG W N， BAI W W， LI J H， et al. Leukemia inhibitory factor is a novel biomarker to predict lymph node and distant metastasis in pancreatic cancer［J］. Int J Cancer， 2021， 148（4）： 1006-1013.
26	ZHANG C， LIU J， WANG J M， et al. The emerging role of leukemia inhibitory factor in cancer and therapy［J］. Pharmacol Ther， 2021， 221： 107754.
27	VAZIRI N， SHARIATI L， JAVANMARD S H. Leukemia inhibitory factor： a main controller of breast cancer［J］. J Biosci， 2020， 45： 143.
28	WANG D， LIU K， YANG Y C， et al. Prognostic value of leukemia inhibitory factor and its receptor in pancreatic adenocarcinoma［J］. Future Oncol， 2020， 16（3）： 4461-4473.
29	LIU B， LU Y， LI J Z， et al. Leukemia inhibitory factor promotes tumor growth and metastasis in human osteosarcoma via activating STAT3［J］. APMIS， 2015， 123（10）： 837-846.
30	LIU Y N， NIU S X， CHEN W Y， et al. Leukemia inhibitory factor promotes castration-resistant prostate cancer and neuroendocrine differentiation by activated ZBTB46［J］. Clin Cancer Res， 2019， 25（13）：4128-4140.
31	PASCUAL-GARCÍA M， BONFILL-TEIXIDOR E， PLANAS-RIGOL E， et al. LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8⁺ T cell tumor-infiltration impairing anti-PD1 therapy［J］. Nat Commun， 2019， 10（1）： 2416.
32	KAMOHARA H， OGAWA M， ISHIKO T， et al. Leukemia inhibitory factor functions as a growth factor in pancreas carcinoma cells： involvement of regulation of LIF and its receptor expression［J］. Int J Oncol， 2007， 30（4）： 977-983.
33	KUPHAL S， WALLNER S， BOSSERHOFF A K. Impact of LIF （leukemia inhibitory factor） expression in malignant melanoma［J］. Exp Mol Pathol， 2013，95（2）： 156-165.
34	LI X Y， YANG Q F， YU H Y， et al. LIF promotes tumorigenesis and metastasis of breast cancer through the AKT-mTOR pathway［J］. Oncotarget， 2014， 5（3）： 788-801.
35	WANG J B， XIE C M， PAN S H， et al. N-myc downstream-regulated gene 2 inhibits human cholangiocarcinoma progression and is regulated by leukemia inhibitory factor/microRNA-181c negative feedback pathway［J］. Hepatology， 2016， 64（5）： 1606-1622.

[1]	Zhuangzhi WU,Xiaoning HE,Siqi CHEN. Inhibitory effect of miR-124-3p on proliferation and invasion of oral squamous cell carcinoma cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 718-727.
[2]	Jing WANG,Chang XU,Yilan SONG,Chongyang WANG,Jingzhi JIANG,Liangchang LI,Guanghai YAN,Liming SU. Alleviation of esculentoside A on airway inflammation of asthmatic mice and its effect on expression levels of IL-6 and STAT3 in lung tissue [J]. Journal of Jilin University(Medicine Edition), 2022, 48(2): 299-307.
[3]	Juan CAO,Weibo LI,Xiu GUO,Bo LI,Chunling DONG. Effects of targeted silencing of heat shock protein 27 on invasion and migration of oral squamous cell carcinoma CAL27 cells and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2021, 47(4): 971-977.
[4]	Yanyan FAN,Na WANG,Wen XU,Yajie GE,Lina WANG. Preventive effect of fresh amniotic membrane on intrauterine adhesion and its improvement effect on endometrial receptivity in rabbits [J]. Journal of Jilin University(Medicine Edition), 2021, 47(4): 990-998.
[5]	LIU Luyao, ZHANG Wenwen, CAO Juan, LI Weibo, SUN Hongchen, LI Bo. Effect of curcumin on gene expressions of cytokines secreted by tumor-associated macrophages [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 930-936.
[6]	BAN Shuofeng, ZHANG Shichen, SUN Yidan, WANG Jing, HAN Bing. Expressions of heterogeneous nuclear ribonucleoprotein D and epithelial cell adhesion molecule in oral squamous cell carcinoma tissue and their significances [J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 589-593.
[7]	DENG Ying, ZHU Yuzhen, WU Kefeng, ZHENG Xuebao, YE Hua. Effect of lncRNA H19 up-regulation mediated by IL-6/ STAT3 pathway in pathogenesis of ulcerative colitis-associated colorectal cancer in mice [J]. Journal of Jilin University(Medicine Edition), 2020, 46(01): 20-25.
[8]	LYU Yanan, FAN Wei, HU Zheng. Construction of immunodeficient mouse models with long-term expression of human cytokines based on piggyBac transposon system [J]. Journal of Jilin University(Medicine Edition), 2020, 46(01): 176-181.
[9]	ZHAO Miao, WANG Yi, ZHANG Ying, FENG Yumei, CAO Yawen, JIANG Haisen, LI Wei. Improvement effect of curcumin on cognitive function in mice with sleep deprivation and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1373-1378.
[10]	WANG Ye, WANG Hong, ZHANG Shujian, JI Huayi, JIN Zhengyong. Intervention effect of glutamine on hyperoxia lung injury in neonatal rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 747-751.
[11]	YAO Hongyue, LIU Xinyu, LIU Wanzhu. Protective effect of panaxdiol on acute liver damage of rats induced by carbon tetrachloride and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 479-483.
[12]	GUO Jia, SONG Siqi, WANG Keyu. Changes of inflammatory cytokine levels and P38MAPK/P65NF-κB signaling pathway in cerebral cortex tissue of mice with heat stroke [J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 566-571.
[13]	HEI Lijuan, WANG Juanhong, WEI Wei, FANG Hangrong, DUAN Ying, MA Shifen, XU Junrong. Detection of interleukin-6 levels in serum and thyroid tissue of patients with papillary thyroid carcinoma and their significances [J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 601-605.
[14]	HE Qi, SHI Hua, DAN Yang, SHAO Suju. Effects of Jiawei Danshen Yin on cardiac function and serum inflammatory factors IL-6 and TNF-α in rats with chronic heart failure [J]. Journal of Jilin University(Medicine Edition), 2019, 45(01): 63-68.
[15]	YANG Wei, LI Mingcheng. Analysis on gene profile of over-expression WWOX gene in human oral squamous cell carcinoma Tca8113 cells and tumor inhibition mechanism of WWOX gene [J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1169-1173.

Promotion effect of tumor associated macrophages-derived IL-6 on invasion and migration of oral squamous cell carcinoma Cal27 cells by upregulating LIF expression in tumor cells

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 35

Related Articles 15

Metrics

Comments

Recommended 0