A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis

Yuan HE; Juncheng GUO; Zhibin YE; Xiaohu WANG; Haonan LI; Jingbiao HUANG

doi:10.19401/j.cnki.1007-3639.2025.06.004

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A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis

Article | 更新时间：2025-12-31

- A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis
- China Oncology Vol. 35, Issue 6, Pages: 555-562(2025)
- 作者机构：
  
  1. 长治医学院附属和平医院胃肠外科，山西长治 046000
  2. 长治医学院第一临床学院，山西长治 046000
  3. 河北省人民医院胃肠外科，河北石家庄 050051
- 作者简介：
  
  HE Yuan
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2025.06.004
  CLC： R735.9
- Received：20 December 2024，
  
  Revised：2025-04-03，
  
  Published：30 June 2025
- 稿件说明：
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Yuan HE, Juncheng GUO, Zhibin YE, et al. A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis[J]. China Oncology, 2025, 35(6): 555-562.
DOI：

Yuan HE, Juncheng GUO, Zhibin YE, et al. A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis[J]. China Oncology, 2025, 35(6): 555-562. DOI： 10.19401/j.cnki.1007-3639.2025.06.004.

摘要

背景与目的：

丝氨酸蛋白酶抑制剂（serine protease inhibitor，SERPIN）的家族成员可以通过调节肿瘤细胞的凋亡、侵袭、转移和血管生成等过程，影响肿瘤的发生、发展和预后，但其在胰腺癌中的作用机制尚不清楚。本研究旨探讨丝氨酸蛋白酶抑制剂A3蛋白（serine protease inhibitors A3，SERPINA3）高表达与胰腺癌细胞的增殖、凋亡、迁移及吉西他滨耐药之间的关系及机制。

方法：

通过实时定量反转录聚合酶链反应（real-time quantitative reverse transcription polymerase chain reaction，qRT-PCR）分析正常胰腺导管上皮细胞系hTERT-HPNE与胰腺癌细胞系SW1990、Capan-1、PANC-1、AsPC-1中SERPINA3的表达水平；构建吉西他滨耐药胰腺癌细胞系PANC-1/R、AsPC-1/R，采用qRT-PCR实验、细胞计数试剂盒-8（cell counting kit-8，CCK-8）测定耐药及亲代敏感细胞系中SERPINA3表达水平及其对吉西他滨的药物敏感性差异。采用小干扰RNA（small interfering RNA，siRNA）构建SERPINA3敲低细胞系si-SERPINA，以siRNA negative control构建阴性对照组si-SERPINA#NC；利用丙二醛（malondialdehyde，MDA）、CCK-8、EdU细胞增殖实验、transwell迁移、matrigel侵袭、划痕和细胞凋亡实验，分别检测si-SERPINA组和si-SERPINA#NC组对吉西他滨耐药胰腺癌细胞的脂质过氧化水平、增殖、迁移、侵袭、修复能力和细胞凋亡的影响。

结果：

与正常胰腺导管上皮细胞hTERT-HPNE相比，SERPINA3在多种胰腺癌细胞系中的表达水平显著升高（

0.05）。相较于亲本细胞PANC-1和AsPC-1，其耐药衍生细胞PANC-1/R和AsPC-1/R细胞株中SERPINA3的mRNA表达和蛋白水平均显著升高（

0.001）。当

SERPINA3

在PANC-1/R和AsPC-1/R细胞中被敲低后，细胞在不同浓度吉西他滨作用下的存活率下降，并且在MDA检测中显示出脂质过氧化水平增加（

0.001）。此外，敲低

SERPINA3

的PANC-1/R和AsPC-1/R细胞系株的增殖率、迁移/侵袭细胞数量以及划痕实验的愈合率均显著降低（

0.01），流式细胞术检测到凋亡细胞的数量增加（

0.05）。这些结果表明，敲低SERPINA3能够抑制吉西他滨耐药胰腺癌细胞的增殖、迁移、侵袭和修复能力

，并促进这些耐药细胞的凋亡。

结论：

SERPINA3在多种胰腺癌细胞中的呈高表达，SERPINA3高表达促进胰腺癌恶性进展和耐药，干扰SERPINA3表达可促进铁死亡并增强吉西他滨耐药胰腺癌细胞对药物的敏感性。

Abstract

Background and purpose:

Members of the serine protease inhibitor (SERPIN) family can influence tumorigenesis

progression

and prognosis by modulating processes such as apoptosis

invasion

metastasis

and angiogenesis in tumor cells. However

their role in pancreatic cancer remains unclear. This study aimed to investigate the impact of high expression of serine protease inhibitor A3 (SERPINA3) on the proliferation

apoptosis

migration

and chemoresistance of pancreatic cancer cells and its mechanism.

Methods:

This study analyzed the SERPINA3 expression levels in the normal pancreatic ductal epithelial cell line hTERT-HPNE and pancreatic cancer cell lines SW1990

Capan-1

PANC-1

and ASPC-1 by real-time reverse transcription quantitative polymerase chain reaction (qRT-PCR). We established gemcitabine-resistant pancreatic cancer cell lines PANC-1/R and ASPC-1/R

and used qRT-PCR assay and cell counting kit-8 (CCK-8) to determine the SERPINA3 expression levels in the constructed resistant cell lines and their parental sensitive cell lines

as well as the differences in their chemosensitivity to gemcitabine. We constructed the

SERPINA3

-knockdown cell line si-SERPINA with siRNA

and the negative control group si-SERPINA#NC with siRNA negative control. We used MDA assay

CCK-8 assay

EdU cell proliferation assay

transwell migration assay

matrigel invasion assay

scratch assay

and apoptotic assay to respectively detect the lipid oxidation levels

proliferation

migration

invasion

wound-healing ability

and the influence on apoptosis of the gemcitabine-resistant pancreatic cancer cells in the si-SERPINA group and the si-SERPINA#NC group.

Results:

Compared with normal pancreatic ductal epithelial cells hTERT-HPNE

the expression level of SERPINA3 in

various pancreatic cancer cell lines was significantly increased (

0.05). mRNA and protein expression levels of SERPINA3 in PANC-1/R and ASPC-1/R were significantly increased compared with those in parent cells (

0.001). When

SERPINA3

was knocked down in PANC-1/R and ASPC-1/R cells

the survival rate of the cells under different concentrations of gemcitabine chemotherapy decreased

and MDA detected that the lipid oxidation level was increased (

0.001). In addition

the proliferation rate of PANC-1/R and ASPC-1/R cell lines with

SERPINA3

knockout

the number of migrating/invading cells and the healing rate of scratch test were significantly decreased (

0.01)

and flow cytometry demonstrated that the number of apoptotic cells was increased (

0.05). These results suggest that

SERPINA3

knockdown can inhibit the proliferation

migration

invasion and wound healing ability of gemcitabine-resistant pancreatic cancer cells

and promote the apoptosis of these resistant cells.

Conclusion:

SERPINA3 overexpression was found in various pancreatic cancer cells. SERPINA3 overexpression promoted malignant progression and chemotherapy resistance of pancreatic cancer

and interference with SERPINA3 expression promoted ferroptosis and enhanced chemotherapy sensitivity of gemcitabine-resistant pancreatic cancer cells.

关键词

Keywords

references

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