NFATc3在恶性黑素瘤中的表达及其对细胞存活和转移的影响

张婉琪; 沈潇潇; 李红丽; 鲁元刚

doi:10.16016/j.1000-5404.202101108

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NFATc3在恶性黑素瘤中的表达及其对细胞存活和转移的影响

基础医学 | 更新时间：2026-01-09

- NFATc3在恶性黑素瘤中的表达及其对细胞存活和转移的影响
- Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis
- 第三军医大学学报 2021年43卷第11期页码：1080-1088
- 作者机构：
  
  1.400042 重庆，陆军军医大学(第三军医大学)大坪医院整形美容科
  2.400038 重庆，陆军军医大学(第三军医大学)基础医学院教学实验中心
- 作者简介：
  
  [鲁元刚，E-mail: skin515@163.com LU Yuangang, E-mail: skin515@163.com
- 基金信息：
  
  陆军军医大学科技创新能力提升专项(2019XQY01)
- DOI：10.16016/j.1000-5404.202101108
  中图分类号： R394.3;R730.23;R739.5
- 收稿：2021-01-12，
  
  修回：2021-02-08，
  
  纸质出版：2021-06-15
- 稿件说明：
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张婉琪, 沈潇潇, 李红丽, 等. NFATc3在恶性黑素瘤中的表达及其对细胞存活和转移的影响[J]. 第三军医大学学报, 2021,43(11):1080-1088.

Wanqi ZHANG, Xiaoxiao SHEN, Hongli LI, et al. Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis[J]. Journal of Third Military Medical University, 2021, 43(11): 1080-1088.
张婉琪, 沈潇潇, 李红丽, 等. NFATc3在恶性黑素瘤中的表达及其对细胞存活和转移的影响[J]. 第三军医大学学报, 2021,43(11):1080-1088. DOI： 10.16016/j.1000-5404.202101108.

Wanqi ZHANG, Xiaoxiao SHEN, Hongli LI, et al. Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis[J]. Journal of Third Military Medical University, 2021, 43(11): 1080-1088. DOI： 10.16016/j.1000-5404.202101108.

摘要

目的

探讨NFATc3在不同恶性程度的恶性黑素瘤(malignant melanoma，MM)组织及细胞中的表达及意义。

方法

收集转移(11例)和未转移(20例)MM组织及癌旁组织(3例)石蜡切片；对比不同恶性度MM细胞系，运用免疫组化染色、Crispr/Cas9敲减技术并转染细胞，通过qPCR、Western blot、CCK-8及流式细胞学等方法观察NFATc3表达对MM细胞活性、增殖凋亡与侵袭迁移的影响。

结果

① NFATc3在转移性MM组织中呈高表达，在癌旁组织中几乎无表达(

＜0.01)；在转移性MM细胞Mel-RM中，NFATc3 mRNA和蛋白水平较原位来源MM细胞A375显著上调(

＜0.01)；②敲减NFATc3后，敲减组的细胞存活率显著下降(

0.01)，而Caspase-3阳性荧光细胞比例则显著升高(

0.01)；流式细胞检测及TUNEL染色结果显示，敲减组中凋亡细胞比例增加(

0.01)；Western blot检测发现敲减组细胞中活化的Caspase-3(17×10

)和Caspase-8(18×10

)蛋白表达增高；③划痕实验及Transwell检测均表明敲减组中侵袭和迁移的细胞数目显著减少(

0.01)。

结论

NFATc3在转移性MM中表达上调，且其表达水平与恶性程度相关。下调NFATc3能减少MM细胞存活、促进细胞凋亡并抑制细胞转移能力。

Abstract

Objective

To investigate the expression and significance of NFATc3 in different malignant melanoma (MM) tissues and cells.

Methods

The metastatic (11 cases) and non-metastatic (20 cases) MM tissues and adjacent tissues (3 cases) were collected in paraffin sections. MM cell lines with different malignancy were compared and transfected with Crispr/Cas9 knock-down plasmid. The effects of NFATc3 expression on the proliferation

apoptosis and invasion of MM cells were observed by immunohistochemical staining

qPCR

Western blotting

CCK-8 assay and flow cytometry.

Results

① NFATc3 was highly expressed in MM tissues

but hardly seen in the adjacent tissues (

0.01). Its expression at mRNA and protein levels was obviously up-regulated in the Mel-RM cells when compared with the A375 cells (

0.01). ②After NFATc3 knock-down

the cell survival rate of NFATc3 knock-down group was significantly decreased (

0.05). The proportion of Caspase-3 positive fluorescent cells was significantly increased (

0.01). Flow cytometry and TUNEL staining showed an increased proportion of apoptotic cells in the knock-down group (

0.05)

and further Western blot assay confirmed that the expression of activated Caspase-3 (17×10

) and Caspase-8 (18×10

) proteins were increased. ③The number of invaded and migrated cells in the knock-down group were significantly reduced by wound healing test and Transwell test (

0.01).

Conclusion

NFATc3 is highly expressed in metastatic MM

and its expression level is correlated to the malignancy. NFATc3 knockdown can induce the apoptosis and inhibit the invasion and migration abilities of MM cells.

关键词

Keywords

references

WHITEMAN D C, GREEN A C, OLSEN C M. The growing burden of invasive melanoma: projections of incidence rates and numbers of new cases in six susceptible populations through 2031[J]. J Invest Dermatol, 2016, 136(6): 1161-1171.

DAVIS L E, SHALIN S C, TACKETT A J. Current state of melanoma diagnosis and treatment[J]. Cancer Biol Ther, 2019, 20(11): 1366-1379.

NAMIKAWA K, YAMAZAKI N. Targeted therapy and immunotherapy for melanoma in Japan[J]. Curr Treat Options Oncol, 2019, 20(1): 7.

SOSMAN J A, KIM K B, SCHUCHTER L, et al . Survival in BRAF V600-mutant advanced melanoma treated with vemurafenib[J ] . N Engl J Med, 2012, 366(8): 707-714.

LAN X, ZHAO J, SONG C, et al . TRPM8 facilitates proliferation and immune evasion of esophageal cancer cells[J ] . Biosci Rep, 2019, 39(10): BSR20191878.

LIU X, WANG T, WANG Y, et al . Orai1 is critical for Notch-driven aggressiveness under hypoxic conditions in triple-negative breast cancers[J ] . Biochim Biophys Acta Mol Basis Dis, 2018, 1864(4 pt a): 975-986.

URSO K, FERNÁNDEZ A, VELASCO P, et al . NFATc3 controls tumour growth by regulating proliferation and migration of human astroglioma cells[J ] . Sci Rep, 2019, 9(1): 9361.

XIAO T, CHEN W C, WANG S F, et al . Tacrolimus and ascomycin inhibit melanoma cell growth, migration and invasion via targeting nuclear factor of activated T-cell 3[J ] . Melanoma Res, 2020, 30(4): 325-335.

MOGNOL G P, CARNEIRO F R, ROBBS B K, et al . Cell cycle and apoptosis regulation by NFAT transcription factors: new roles for an old player[J ] . Cell Death Dis, 2016, 7: e2199.

WANG G, WANG Y Z, YU Y, et al . The critical role of calcineurin/NFAT (C/N) pathways and effective antitumorprospect for colorectal cancers[J ] . J Cell Biochem, 2019, 120(12): 19254-19273.

SHOU J, JING J, XIE J, et al . Nuclear factor of activated T cells in cancer d evelopment and treatment[J ] . Cancer Lett, 2015, 361(2): 174-184.

BUCHER P, ERDMANN T, GRONDONA P, et al . Targeting chronic NFAT activation with calcineurin inhibitors in diffuse large B-cell lymphoma[J ] . Blood, 2020, 135(2): 121-132.

XIN B, JI K Q, LIU Y S, et al . Higher expression of calcineurin predicts poor prognosis in unique subtype of ovarian cancer[J ] . J Ovarian Res, 2019, 12(1): 75.

LIU X, WAN X, KAN H, et al . Hypoxia-induced upregulation of Orai1 drives colon cancer invasiveness and angiogenesis[J ] . Eur J Pharmacol, 2018, 832: 1-10.

CARNEIRO B A, EL-DEIRY W S. Targeting apoptosis in cancer therapy[J]. Nat Rev Clin Oncol, 2020, 17(7): 395-417.

ZHAO B, XIE J, ZHOU X, et al . YAP activation in melanoma contributes to anoikis resistance and metastasis[J ] . Exp Biol Med (Maywood), 2020: 1535370220977101.

BUTTERICK T A, IGBAVBOA U, ECKERT G P, et al . Simvastatin stimulates production of the antiapoptotic protein Bcl-2 via endothelin-1 and NFATc3 in SH-SY5Y cells[J ] . Mol Neurobiol, 2010, 41(2/3): 384-391..

NERIA F, DEL CARMEN SERRANO-PEREZ M, VELASCO P, et al . NFATc3 promotes Ca 2+ -dependent MMP3 expression in astroglial cells[J ] . Glia, 2013, 61(7): 1052-1066..

SCHEAU C, BADARAU I A, COSTACHE R, et al . The role of matrix metalloproteinases in the epithelial-mesenchymal transition of hepatocellular carcinoma[J ] . Anal Cell Pathol (Amst), 2019, 2019: 9423907.

袁媛, 费乐学, 刘岩, 等. 基质金属蛋白酶-3与细胞外基质金属蛋白酶诱导因子在乳腺癌演变过程中的表达及意义[J]. 实用医学杂志, 2008, 24(1): 10-13.

YUAN Y, FEI L X, LIU Y, et al . Expressions of matrix metalloproteinase-3 and extracellular matrix metallo-proteinase inducer in the carcinogenesis of breast cancer and theirs significances[J ] . J Pract Med, 2008, 24(1): 10-13.

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