Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis

Wanqi ZHANG; Xiaoxiao SHEN; Hongli LI; Yuangang LU

doi:10.16016/j.1000-5404.202101108

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Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis

Basic Medicine | 更新时间：2026-01-09

- Expression of NFATc3 in malignant melanoma and its effects on cell survival and metastasis
- Journal of Third Military Medical University Vol. 43, Issue 11, Pages: 1080-1088(2021)
- 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：
- 基金信息：
  
  the Promotion Project of Scientific and Technological Innovation Abilities of Army Medical University(2019XQY01)
- DOI：10.16016/j.1000-5404.202101108
  CLC： R394.3;R730.23;R739.5
- Received：12 January 2021，
  
  Revised：2021-02-08，
  
  Published：15 June 2021
- 稿件说明：
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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：

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

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