Research on the role of ACE2 in drug resistance of colorectal cancer and its predictive vaule

doi:10.11958/20260224

Abstract

Abstract:

Objective To investigate the role of angiotensin-converting enzyme 2 (ACE2) in chemoresistance in colorectal cancer (CRC) and evaluate its potential as a predictive biomarker. Methods Serum samples were collected from patients with colorectal cancer and healthy controls, and ACE2 protein levels were measured by enzyme-linked immunosorbent assay (ELISA). Baseline and drug-induced (cisplatin, 5-fluorouracil and paclitaxel) ACE2 expression in CRC cell lines were detected by Western blot assay and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Small interfering RNAs (siRNAs) and lentiviral systems were employed to construct ACE2 knockdown and stable overexpression cell models. The effect of ACE2 expression on chemoresistance was evaluated by calculating the half-maximal inhibitory concentration (IC₅₀) for cisplatin, 5-fluorouracil and paclitaxel. Cell proliferation and migration abilities were assessed using the Cell Counting Kit-8 (CCK-8) assay, colony formation assay and Transwell assay, respectively. The association between ACE2 expression and treatment response or prognosis in CRC patients was analyzed using clinical specimen and public database. Results Clinical data revealed that serum ACE2 levels were higher in the healthy controls than those in colorectal cancer patients (P<0.05). Serum ACE2 levels were higher in the chemotherapy-resistant group than those in the chemotherapy-sensitive group（P<0.05). Exposure to chemotherapeutic agents (cisplatin, 5-fluorouracil, and paclitaxel) upregulated ACE2 expression in colorectal cancer cells（P<0.05). Knockdown of ACE2 reduced the IC₅₀ value of these drugs, and significantly inhibited cell proliferation and migration（P<0.05). Bioinformatics analysis showed that ACE2 expression was correlated with multiple drug resistance-related genes. Moreover, among colorectal cancer patients with low KRAS expression, high ACE2 expression was associated with poorer prognosis（P<0.05). Conclusion ACE2 expression is positively correlated with malignant phenotypes (proliferation and migration) and chemoresistance in CRC cells. Elevated ACE2 levels in patients after chemotherapy can serve as a potential biomarker for predicting chemoresistance.

Key words: angiotensin-converting enzyme 2, colorectal neoplasms, cell proliferation, cell movement, prognosis, chemotherapy resistance

CLC Number:

R735.3

Figures/Tables 21

References 19

[1]	BADER M, STECKELINGS U M, ALENINA N, et al. Alternative renin-angiotensin system[J]. Hypertension, 2024, 81(5):964-976. doi:10.1161/HYPERTENSIONAHA.123.21364.
[2]	TANG X, LU L, LI X, et al. Bridging cancer and COVID-19:The complex interplay of ACE2 and TMPRSS2[J]. Cancer Med, 2025, 14(7):e70829. doi:10.1002/cam4.70829.
[3]	IWAI M, HORIUCHI M. Devil and angel in the renin-angiotensin system:ACE-angiotensin II-AT1 receptor axis vs. ACE2-angiotensin-(1-7)-Mas receptor axis[J]. Hypertens Res, 2009, 32(7):533-536. doi:10.1038/hr.2009.74.
[4]	TIKELLIS C, BERNARDI S, BURNS W C. Angiotensin-converting enzyme 2 is a key modulator of the renin-angiotensin system in cardiovascular and renal disease[J]. Curr Opin Nephrol Hypertens, 2011, 20(1):62-68. doi:10.1097/MNH.0b013e328341164a.
[5]	BEYERSTEDT S, CASARO E B, RANGEL É B. COVID-19:angiotensin-converting enzyme 2(ACE2)expression and tissue susceptibility to SARS-CoV-2 infection[J]. Eur J Clin Microbiol Infect Dis, 2021, 40(5):905-919. doi:10.1007/s10096-020-04138-6.
[6]	XI Y, XU P. Global colorectal cancer burden in 2020 and projections to 2040[J]. Transl Oncol, 2021, 14(10):101174. doi:10.1016/j.tranon.2021.101174.
[7]	LI Q, ODURO P K, GUO R, et al. Oncolytic viruses:Immunotherapy drugs for gastrointestinal malignant tumors[J]. Front Cell Infect Microbiol, 2022, 12:921534. doi:10.3389/fcimb.2022.921534.
[8]	ZHANG Q, LU S, LI T, et al. ACE2 inhibits breast cancer angiogenesis via suppressing the VEGFa/VEGFR2/ERK pathway[J]. J Exp Clin Cancer Res, 2019, 38(1):173. doi:10.1186/s13046-019-1156-5.
[9]	RAGO V, BOSSIO S, LOFARO D, et al. New insights into the link between SARS-CoV-2 infection and renal cancer[J]. Life(Basel), 2023, 14(1):52. doi:10.3390/life14010052.
[10]	HASSANI B, ATTAR Z, FIROUZABADI N. The renin-angiotensin-aldosterone system(RAAS)signaling pathways and cancer:foes versus allies[J]. Cancer Cell Int, 2023, 23(1):254. doi:10.1186/s12935-023-03080-9.
[11]	AHMADI M, PASHANGZADEH S, MOUSAVI P, et al. ACE2 correlates with immune infiltrates in colon adenocarcinoma:Implication for COVID-19[J]. Int Immunopharmacol, 2021, 95:107568. doi:10.1016/j.intimp.2021.107568.
[12]	CHOONG O K, JAKOBSSON R, BERGDAHL A G, et al. SARS-CoV-2 replicates and displays oncolytic properties in clear cell and papillary renal cell carcinoma[J]. PLoS One, 2023, 18(1):e0279578. doi:10.1371/journal.pone.0279578.
[13]	KUNVARIYA A D, DAVE S A, MODI Z J, et al. Exploration of multifaceted molecular mechanism of angiotensin-converting enzyme 2(ACE2)in pathogenesis of various diseases[J]. Heliyon, 2023, 9(5):e15644. doi:10.1016/j.heliyon.2023.e15644.
[14]	DONG F, LI H, LIU L, et al. ACE2 negatively regulates the Warburg effect and suppresses hepatocellular carcinoma progression via reducing ROS-HIF1α activity[J]. Int J Biol Sci, 2023, 19(8):2613-2629. doi:10.7150/ijbs.81498.
[15]	WU D, DING Y, WANG T, et al. Significance of tumor-infiltrating immune cells in the prognosis of colon cancer[J]. Onco Targets Ther, 2020, 13:4581-4589. doi:10.2147/OTT.S250416.
[16]	ZUO X, REN S, ZHANG H, et al. Chemotherapy induces ACE2 expression in breast cancer via the ROS-AKT-HIF-1α signaling pathway:a potential prognostic marker for breast cancer patients receiving chemotherapy[J]. J Transl Med, 2022, 20(1):509. doi:10.1186/s12967-022-03716-w.
[17]	ZHANG S, KAPOOR S, TRIPATHI C, et al. Targeting ACE2-BRD4 crosstalk in colorectal cancer and the deregulation of DNA repair and apoptosis[J]. NPJ Precis Oncol, 2023, 7(1):20. doi:10.1038/s41698-023-00361-4.
[18]	KIM D E, OH H J, KIM H J, et al. Synergistic two-step inhibition approach using a combination of trametinib and onvansertib in KRAS and TP53-mutated colorectal adenocarcinoma[J]. Biomed Pharmacother, 2025, 182:117796. doi:10.1016/j.biopha.2024.117796.
[19]	HUANG L, GUO Z, WANG F, et al. KRAS mutation:from undruggable to druggable in cancer[J]. Signal Transduct Target Ther, 2021, 6(1):386. doi:10.1038/s41392-021-00780-4.

基因名称	引物序列（5'→3'）	产物大小/bp
ACE2	上游：GTGCACAAAGGTGACAATGG	160
ACE2	下游：GGCTGCAGAAAGTGACATGA	160
β-actin	上游：CTGGGTGTTGAAGGTCTC	216
β-actin	下游：CAGAGCAAGAGAGGCATCC	216

基因名称	引物序列（5'→3'）	产物大小/bp
ACE2	上游：GTGCACAAAGGTGACAATGG	160
ACE2	下游：GGCTGCAGAAAGTGACATGA	160
β-actin	上游：CTGGGTGTTGAAGGTCTC	216
β-actin	下游：CAGAGCAAGAGAGGCATCC	216

引物名称	序列（5'→3'）
siACE2#1	上游：GCGAGUGGCUAAUUUGAAATT
siACE2#1	下游：UUUCAAAUUAGCCACUCGCTT
siACE2#2	上游：GGACAAGUUUAACCACGAATT
siACE2#2	下游：UUCGUGGUUAAACUUGUCCTT

引物名称	序列（5'→3'）
siACE2#1	上游：GCGAGUGGCUAAUUUGAAATT
siACE2#1	下游：UUUCAAAUUAGCCACUCGCTT
siACE2#2	上游：GGACAAGUUUAACCACGAATT
siACE2#2	下游：UUCGUGGUUAAACUUGUCCTT

组别		n	年龄/岁			化疗反应				手术		复发		转移
组别		n	年龄/岁			耐药		敏感		手术		复发		转移
ACE2低表达组		47	64.00（59.00，72.00）			6（12.80）		41（87.20）		29（61.70）		1（2.10）		18（38.30）
ACE2高表达组		46	63.50（56.75，67.50）			38（82.60）		8（14.70）		31（67.40）		0（0.00）		17（37.00）
Z或χ2			1.181			45.492^＊＊				0.329		0.989		0.018
组别	肿瘤分级				T分期				N分期				M分期
组别	Ⅰ级			Ⅱ—Ⅲ级	T1—T2		T3—T4		N0		N1—N3		M0		MX
ACE2低表达组	12（25.50）			35（74.50）	13（27.70）		34（72.30）		29（61.70）		18（38.30）		28（59.60）		19（40.40）
ACE2高表达组	14（30.40）			32（69.60）	14（30.40）		32（69.60）		28（60.90）		18（39.10）		28（60.90）		18（39.10）
χ2	0.277				0.087				0.007				0.016