敲低TMCO1对肝癌细胞增殖和侵袭的影响

doi:10.3969/j.issn.1000-1565.2024.06.010

摘要/Abstract

摘要： 为探讨跨膜和卷曲螺旋结构域 1(transmembrane and coiled-coil domains 1,TMCO1)对肝癌细胞的增殖和侵袭的影响,构建肝癌细胞HuH-7和Li-7的过表达和敲低TMCO1细胞模型,采用Transwell、细胞克隆、线粒体通透性转换孔(mPTP)探针、线粒体膜电位(JC-1)探针、Actin-Tracker Green-488染色、 Western blot和免疫共沉淀实验检测敲低TMCO1对肝癌细胞增殖、侵袭、线粒体功能、骨架重塑及VDAC1表达的影响.结果显示:敲低TMCO1可抑制肝癌细胞的增殖、侵袭,影响线粒体功能和骨架重塑,并下调VDAC1的表达.

关键词: 肝癌细胞, TMCO1, VDAC1, mPTP, 增殖

Abstract: To explore the effects of transmembrane and coiled-coil domains 1(TMCO1)knockdown on the proliferation and invasion of hepatocellular carcinoma cells, overexpression and knockdown cell models of TMCO1 in hepatocellular carcinoma cell lines HuH-7 and Li-7 were constructed. Transwell, colony formation, mitochondrial permeability transition pore(mPTP)probes, mitochondrial membrane potential(JC-1)probes, Actin-Tracker Green-488 staining, Western blotting and co-immunoprecipitation tests were used to detect the effects of TMCO1 knockdown on hepatocellular carcinoma cells proliferation, invasion, mitochondrial function, cytoskeletal remodeling, and VDAC1 expression. The results indicated that TMCO1 knockdown inhibited the proliferation and invasion of hepatocellular carcinoma cells, affected mitochondrial function and cytoskeletal remodeling, and downregulated VDAC1 expression.

Key words: hepatoma cells, TMCO1, VDAC1, mPTP, proliferation

中图分类号:

R735.7

冷君志,刘迪,王根旺,柳科军,金栋,王琦,惠永峰. 敲低TMCO1对肝癌细胞增殖和侵袭的影响[J]. 河北大学学报(自然科学版), 2024, 44(6): 643-652.

LENG Junzhi, LIU Di, WANG Genwang, LIU Kejun, JIN Dong, WANG Qi, HUI Yongfeng. Effects of TMCO1 knockdown on the proliferation, invasion of hepatocellular carcinoma cells[J]. Journal of Hebei University(Natural Science Edition), 2024, 44(6): 643-652.

参考文献

[1] CHEN W, CHIANG C L, DAWSON L A. Efficacy and safety of radiotherapy for primary liver cancer[J]. Chin Clin Oncol, 2021, 10(1): 9. DOI: 10.21037/cco-20-89.
[2] ZHAO B, LV X C, ZHAO X Q, et al. Tumor-promoting actions of HNRNP A1 in HCC are associated with cell cycle, mitochondrial dynamics, and necroptosis[J]. Int J Mol Sci, 2022, 23(18): 10209. DOI: 10.3390/ijms231810209.
[3] YUAN D Y, CHEN J, HAO Q Y, et al. Methyltransferase-like 3 aggravates HCC development via mediating N6-methyladenosine of ubiquitin-specific protease 7[J]. J Oncol, 2022, 2022: 6167832. DOI: 10.1155/2022/6167832.
[4] NIU M K, YI M, LI N, et al. Advances of targeted therapy for hepatocellular carcinoma[J]. Front Oncol, 2021, 11: 719896. DOI: 10.3389/fonc.2021.719896.
[5] YANG C, ZHANG H L, ZHANG L M, et al. Evolving therapeutic landscape of advanced hepatocellular carcinoma[J]. Nat Rev Gastroenterol Hepatol, 2023, 20(4): 203-222.DOI: 10.1038/s41575-022-00704-9.
[6] BERRIDGE M J, LIPP P, BOOTMAN M D. The versatility and universality of calcium signalling[J]. Nat Rev Mol Cell Biol, 2000, 1(1): 11-21. DOI: 10.1038/35036035.
[7] VAZ C V, RODRIGUES D B, SOCORRO S, et al. Effect of extracellular calcium on regucalcin expression and cell viability in neoplastic and non-neoplastic human prostate cells[J]. Biochim Biophys Acta, 2015, 1853(10 Pt A): 2621-2628. DOI: 10.1016/j.bbamcr.2015.07.006.
[8] WANG Q C, ZHENG Q X, TAN H Y, et al. TMCO1 is an ER Ca(2+)load-activated Ca(2+)channel[J]. Cell, 2016, 165(6): 1454-1466. DOI: 10.1016/j.cell.2016.04.051.
[9] PAREKH A B. Store-operated CRAC channels: function in health and disease[J]. Nat Rev Drug Discov, 2010, 9(5): 399-410.DOI: 10.1038/nrd3136.
[10] SUN Z S, ZHANG H, WANG X, et al. TMCO1 is essential for ovarian follicle development by regulating ER Ca²⁺ store of granulosa cells[J]. Cell Death Differ,2018, 25(9):1686-1701. DOI: 10.1038/s41418-018-0067-x.
[11] XIN B Z, PUFFENBERGER E G, TURBEN S, et al. Homozygous frameshift mutation in TMCO1 causes a syndrome with craniofacial dysmorphism, skeletal anomalies, and mental retardation[J]. Proc Natl Acad Sci USA, 2010, 107(1): 258-263. DOI: 10.1073/pnas.0908457107.
[12] KONDKAR A A, MOUSA A, AZAD T A, et al. Polymorphism rs7555523 in transmembrane and coiled-coil domain 1(TMCO1)is not a risk factor for primary open angle glaucoma in a Saudi cohort[J]. J Negat Results Biomed, 2016, 15(1): 17. DOI: 10.1186/s12952-016-0060-1.
[13] GAO L,YE Z, LIU J H, et al. TMCO1 expression promotes cell proliferation and induces epithelial-mesenchymal transformation in human gliomas[J]. Med Oncol, 2022, 39(5): 90. DOI: 10.1007/s12032-022-01687-y.
[14] YANG C, WANG Y, BAI J Q, et al. Mechanism of transmembrane and coiled-coil domain 1 in the regulation of proliferation and migration of A549 cells[J]. Oncol Lett, 2020, 20(5): 159. DOI: 10.3892/ol.2020.12020.
[15] SIGAL M, LOGAN C Y, KAPALCZYNSKA M, et al. Stromal R-spondin orchestrates gastric epithelial stem cells and gland homeostasis[J]. Nature, 2017, 548(7668): 451-455. DOI: 10.1038/nature23642.
[16] CHEN Y F, CHEN Y T, CHIU W T, et al. Remodeling of calcium signaling in tumor progression[J]. J Biomed Sci, 2013, 20(1): 23. DOI: 10.1186/1423-0127-20-23.
[17] ZHANG C Y, LIU B W, SHENG J X, et al. Potential targets for the treatment of MI: GRP75-mediated Ca²⁺transfer in MAM[J]. Eur J Pharmacol, 2024, 971: 176530. DOI: 10.1016/j.ejphar.2024.176530.
[18] ZHANG F, QI C X, YAO Z P, et al. Identification and validation of a novel necroptosis-related molecular signature to evaluate prognosis and immune features in breast cancer[J]. Apoptosis, 2023, 28(11/12): 1628-1645. DOI: 10.1007/s10495-023-01887-5.
[19] LIAO Y J, HAO Y M, CHEN H, et al. Mitochondrial calcium uniporter protein MCU is involved in oxidative stress-induced cell death[J]. Protein Cell, 2015, 6(6): 434-442. DOI: 10.1007/s13238-015-0144-6.
[20] WO L L, ZHANG X, MA C N, et al. LncRNA HABON promoted liver cancer cells survival under hypoxia by inhibiting mPTP opening[J]. Cell Death Discov, 2022, 8(1): 171. DOI: 10.1038/s41420-022-00917-6.
[21] BASSOT A, CHEN J S,TAKAHASHI-YAMASHIRO K, et al. The endoplasmic reticulum kinase PERK interacts with the oxidoreductase ERO1 to metabolically adapt mitochondria[J]. Cell Rep, 2023, 42(1): 111899. DOI: 10.1016/j.celrep.2022.111899.
[22] 齐顺利,赵团结,于晓倩,等.结肠癌中TMCO1、CALR的表达及对转移的影响[J].临床与实验病理学杂志, 2021, 37(12): 1411-1420. DOI: 10.13315/j.cnki.cjcep.2021.12.002.
[23] AN G, PARK J, SONG J, et al. Relevance of the endoplasmic reticulum-mitochondria axis in cancer diagnosis and therapy[J]. Exp Mol Med, 2024, 56(1): 40-50. DOI: 10.1038/s12276-023-01137-3.
[24] WU N N, WANG L F, WANG L, et al. Site-specific ubiquitination of VDAC1 restricts its oligomerization and mitochondrial DNA release in liver fibrosis[J]. Exp Mol Med, 2023, 55(1): 269-280. DOI: 10.1038/s12276-022-00923-9.
[25] ZHOU Q Q, LIU T F, QIAN W J, et al. HNF4A-BAP31-VDAC1 axis synchronously regulates cell proliferation and ferroptosis in gastric cancer[J]. Cell Death Dis, 2023, 14(6): 356. DOI: 10.1038/s41419-023-05868-z.
[26] HUANG L,WEI B, ZHAO Y R, et al. DYNLT1 promotes mitochondrial metabolism to fuel breast cancer development by inhibiting ubiquitination degradation of VDAC1[J]. Mol Med, 2023, 29(1): 72. DOI: 10.1186/s10020-023-00663-0.
[27] HESLOP K A, BURGER P, KAPPLER C, et al. Small molecules targeting the NADH-binding pocket of VDAC modulate mitochondrial metabolism in hepatocarcinoma cells[J]. Biomed Pharmacother, 2022, 150: 112928. DOI: 10.1016/j.biopha.2022.112928.
[28] FREITAS S, MARTINS R, COSTA M, et al. Hierridin B isolated from a marine cyanobacterium alters VDAC1, mitochondrial activity, and cell cycle genes on HT-29 colon adenocarcinoma cells[J]. Mar Drugs, 2016, 14(9): 158. DOI: 10.3390/md14090158.
[29] ZHOU H, ZHANG Y, HU S Y, et al. Melatonin protects cardiac microvasculature against ischemia/reperfusion injury via suppression of mitochondrial fission-VDAC1-HK2-mPTP-mitophagy axis[J]. J Pineal Res, 2017, 63(1): e12413. DOI: 10.1111/jpi.12413.
[30] JIANG L,WANG H, CHEN G B, et al. WDR26/MIP2 interacts with VDAC1 and regulates VDAC1 expression levels in H9c2 cells[J]. Free Radic Biol Med, 2018, 117: 58-65. DOI: 10.1016/j.freeradbiomed.2017.12.015.
[31] HE Y J, WANG W J, YANG T, et al. The potential role of voltage-dependent anion channel in the treatment of Parkinson’s disease[J]. Oxid Med Cell Longev, 2022, 2022: 4665530. DOI: 10.1155/2022/4665530.
[32] HU T, ZOU H X, LE S Y, et al. Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1[J]. Int J Mol Med, 2023, 52(5): 109. DOI: 10.3892/ijmm.2023.5312.
[33] LI J, QI F Z, SU H S, et al. GRP75-faciliated mitochondria-associated ER membrane(MAM)integrity controls cisplatin-resistance in ovarian cancer patients[J]. Int J Biol Sci, 2022, 18(7): 2914-2931. DOI: 10.7150/ijbs.71571. (