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

doi:10.19401/j.cnki.1007-3639.2025.06.004

Abstract

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 (P<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 (P<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 (P<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 (P<0.01), and flow cytometry demonstrated that the number of apoptotic cells was increased (P<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.

Key words: Pancreatic cancer, SERPINA3, Ferroptosis, Gemcitabine, Drug resistance

CLC Number:

R735.9

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

Figures/Tables 8

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

References 16

[1]	BRAY F, LAVERSANNE M, SUNG H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2024, 74(3): 229-263.
[2]	DIXON S J, OLZMANN J A. The cell biology of ferroptosis[J]. Nat Rev Mol Cell Biol, 2024, 25(6): 424-442.
[3]	CHEN Y M, JIANG Z L, ZHANG C X, et al. 5-Methylcytosine transferase NSUN2 drives NRF2-mediated ferroptosis resistance in non-small cell lung cancer[J]. J Biol Chem, 2024, 300(4): 106793.
[4]	ZHU M N, LAN Z H, PARK J, et al. Regulation of CNS pathology by Serpina3n/SERPINA3: The knowns and the puzzles[J]. Neuropathol Appl Neurobiol, 2024, 50(2): e12980.
[5]	MAWARIBUCHI S, SHIMOMURA O, ODA T, et al. rBC2LCN- reactive SERPINA3 is a glycobiomarker candidate for pancreatic ductal adenocarcinoma[J]. Glycobiology, 2023, 33(4): 342-352.
[6]	GUGENHEIM J, CROVETTO A, PETRUCCIANI N. Neoadjuvant therapy for pancreatic cancer[J]. Updates Surg, 2022, 74(1): 35-42.
[7]	WOOD L D, CANTO M I, JAFFEE E M, et al. Pancreatic cancer: pathogenesis, screening, diagnosis, and treatment[J]. Gastroenterology, 2022, 163(2): 386-402.e1.
[8]	LI Y, GUO L. The versatile role of Serpina3c in physiological and pathological processes: a review of recent s dies[J]. Front Endocrinol (Lausanne), 2023, 14: 1189007.
[9]	SOMAN A, ASHA NAIR S. Unfolding the cascade of SERPINA3: inflammation to cancer[J]. Biochim Biophys Acta Rev Cancer, 2022, 1877(5): 188760.
[10]	YANG H, GONG C, WU Y, et al. LncRNA SNHG1 facilitates colorectal cancer cells metastasis by recruiting HNRNPD protein to stabilize SERPINA3 mRNA[J]. Cancer Lett, 2024, 604: 217217.
[11]	LI G, SHE F F, LIAO C Y, et al. cNEK6 induces gemcitabine resistance by promoting glycolysis in pancreatic ductal adenocarcinoma via the SNRPA/PPA2c/mTORC1 axis[J]. Cell Death Dis, 2024, 15(10): 742.
[12]	DE LAAT V, TOPAL H, SPOTBEEN X, et al. Intrinsic temperature increase drives lipid metabolism towards ferroptosis evasion and chemotherapy resistance in pancreatic cancer[J]. Nat Commun, 2024, 15(1): 8540. doi: 10.1038/s41467-024-52978-z pmid: 39358362
[13]	LI Y, RAN Q, DUAN Q, et al. 7-dehydrocholesterol dictates ferroptosis sensitivity[J]. Nature, 2024, 626(7998): 411-8.
[14]	KIM M J, KIM H S, KANG H W, et al. SLC38A5 modulates ferroptosis to overcome gemcitabine resistance in pancreatic cancer[J]. Cells, 2023, 12(20).
[15]	SUN S, SHEN J, JIANG J, et al. Targeting ferroptosis opens new avenues for the development of novel therapeutics[J]. Signal Transduct Target Ther, 2023, 8(1): 372.
[16]	SHI J F, LIU Y, WANG Y, et al. Targeting ferroptosis, a novel programmed cell death, for the potential of alcohol-related liver disease therapy[J]. Front Pharmacol, 2023, 14: 1194343.