Has_circ_0008717在羊膜间充质干细胞上清促血管生成中的作用研究

口腔生物医学 ›› 2024, Vol. 15 ›› Issue (2): 91-96.

Has_circ_0008717在羊膜间充质干细胞上清促血管生成中的作用研究

翟家彬¹,唐子春²,袁乙钦³,高洋⁴,陆史俊⁵,钱文慧²

1. 南京大学附属口腔医院
2. 苏州口腔医院
3. 苏州大学苏州医学院
4. 苏州大学附属第一医院
5. 苏州卫生职业技术学院附属苏州市华夏口腔医院

收稿日期:2023-12-05 修回日期:2024-01-14 出版日期:2024-04-25 发布日期:2024-04-29
通讯作者: 翟家彬 E-mail:zjb1111@aliyun.com
基金资助:
国家自然科学基金

The function of has_circ_0008717 in pro-angiogenic capacity of conditioned medium from human amnion-derived mesenchymal stem cells

Received:2023-12-05 Revised:2024-01-14 Online:2024-04-25 Published:2024-04-29
Contact: Jia binZhai E-mail:zjb1111@aliyun.com
Supported by:
National Natural Science Foundation of China

摘要/Abstract

摘要： 目的：探究has_circ_0008717在人羊膜间充质干细胞上清（hAMSC-CM）促进人脐静脉内皮细胞（HUVECs）血管生成过程中的作用。方法：通过划痕实验、Transwell实验及成管实验检测hAMSC-CM对HUVECs迁移及成管能力的影响，实时荧光定量RT-PCR筛选上调的circRNA（has_circ_0008717）；小干扰RNA（siRNA）下调HUVECs内has_circ_0008717水平后，实时荧光定量RT-PCR检测血管内皮生长因子A（VEGFA）表达水平变化，划痕实验、Transwell实验及成管实验观察其对hAMSC-CM促进HUVECs迁移及成管的影响。结果：hAMSC-CM可明显促进HUVECs迁移及成管能力（P＜0.05），HUVECs中has_circ_0008717升高最为明显（P＜0.05），而敲低has_circ_0008717可明显抑制hAMSC-CM促进HUVECs迁移、成管和VEGFA的表达（P＜0.05）。结论：hAMSC-CM可通过has_circ_0008717增强HUVECs的成管能力。

关键词: has_circ_0008717, 羊膜间充质干细胞, 血管生成, VEGFA

Abstract: Objective:?To investigate the role of has_circ_0008717 during the process that human amnion-derived mesenchymal stem cells (hAMSCs) promote angiogenesis of human umbilical vein endothelial cells (HUVECs). Methods:?Wound healing assay, Transwell migration and tube formation were used to confirm the pro-angiogenic capacity of conditioned medium from hAMSCs (hAMSC-CM) on HUVECs, qRT-PCR and were used to detect the change of level of the potential pro-angiogenic relative circRNAs and genes in HUVECs after treated with hAMSC-CM, among them, has_circ_0008717 and VEGFA attracted our attention. siRNAs specially targeted has_circ_0008717 were used to down-regulate the level of has_circ_0008717 in HUVECs treated with hAMSC-CM, and then qRT-PCR was used to detect were used to evaluate the change of level of VEGFA caused by down-regulation of has_circ_00087, afterwards,wound healing assay, Transwell migration and tube formation were used to assess the effects of down-regulation of has_circ_0008717 on HUVECs treated with hAMSC-CM. Results: Enhanced migration and tube formation in HUVECs induced by hAMSC-CM were observed (P＜0.05). And has_circ_0008717 been the mostly up-regulated circRNA among several potential pro-angiogenic relative circRNAs was selected for further investigation (P＜0.05). Increased expression of vascular endothelial growth factor A (VEGFA) was observed among a few pro-angiogenic relative genes (P＜0.05). Furthermore, these can be inhibited by silencing has_circ_0008717 in HUVEC treated with hAMSC-CM (P＜0.05). Conclusion: has_circ_0008717 might be involved in the pro-angiogenic role of hAMSC-CM in HUVECs.

Key words: circular RNA ABCB10, hAMSCs, angiogenesis, VEGFA

翟家彬唐子春袁乙钦高洋陆史俊钱文慧. Has_circ_0008717在羊膜间充质干细胞上清促血管生成中的作用研究[J]. 口腔生物医学, 2024, 15(2): 91-96.

参考文献 35

[1]	Zhang L, Jiao G, Ren S, et al. Exosomes from bone marrow mesenchymal stem cells enhance fracture healing through the promotion of osteogenesis and angiogenesis in a rat model of nonunion. Stem Cell Res Ther. 2020;11(1):38.
[2]	Hilal,Ahmad,et al.Dual functional approaches for osteogenesis coupled angiogenesis in bone tissue engineering[J].Materials Science & Engineering C, 2019.DOI:10.1016/j.msec.2019.109761.
[3]	Zhang J , Pan J , Jing W .Motivating role of type H vessels in bone regeneration[J].Cell Proliferation, 2020, 53(9).
[4]	Mazini L , Rochette L , Admou B ,et al.Hopes and Limits of Adipose-Derived Stem Cells (ADSCs) and Mesenchymal Stem Cells (MSCs) in Wound Healing.[J].Int J Mol Sci, 2020(4).
[5]	Guillamat-Prats R. The Role of MSC in Wound Healing, Scarring and Regeneration. Cells. 2021;10(7):1729.
[6]	Santos R D A , Asensi K D , Barros J H O D ,et al.Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow[J].International Journal of Molecular Sciences, 2020, 21(24):9563.
[7]	Zuo S, Jones WK, Li H, et al. Paracrine effect of Wnt11-overexpressing mesenchymal stem cells on ischemic injury. Stem Cells Dev. 2012;21(4):598-608.
[8]	Brown C , Mckee C , Bakshi S ,et al.Mesenchymal stem cells: Cell therapy and regeneration potential[J].Journal of Tissue Engineering and Regenerative Medicine, 2019, 13(9).
[9]	Chu,Phuong,Tien,et al.An Update on the Progress of Isolation, Culture, Storage, and Clinical Application of Human Bone Marrow Mesenchymal Stem/Stromal Cells[J].International Journal of Molecular Sciences, 2020, 21(3):708.
[10]	Seo M S , Kang K K , Oh S K ,et al.Isolation and Characterization of Feline Wharton's Jelly-Derived Mesenchymal Stem Cells[J]. 2020.
[11]	Ling L , Feng X , Wei T ,et al.Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism[J].Stem Cell Research & Therapy, 2019, 10(1).
[12]	Liu Q W , Huang Q M , Wu H Y ,et al.Characteristics and Therapeutic Potential of Human Amnion-Derived Stem Cells[J].International Journal of Molecular Sciences, 2021, 22(2):970.
[13]	Jiang F, Ma J, Liang Y, Niu Y, Chen N, Shen M. Amniotic Mesenchymal Stem Cells Can Enhance Angiogenic Capacity via MMPs In Vitro and In Vivo. Biomed Res Int. 2015;2015:324014.
[14]	Li W ,Jia㏎iang Liu, Chen M ,et al.Circular RNA in cancer development and immune regulation[J].Journal of Cellular and Molecular Medicine, 2020(1).
[15]	Kristensen L S , Hansen T B , Ven M T ,et al.Circular RNAs in cancer: opportunities and challenges in the field[J].Oncogene, 2018.
[16]	Misir S, Wu N, Yang BB. Specific expression and functions of circular RNAs. Cell Death Differ. 2022;29(3):481-491.
[17]	He Q, Zhao L, Liu Y, et al. circ-SHKBP1 Regulates the Angiogenesis of U87 Glioma-Exposed Endothelial Cells through miR-544a/FOXP1 and miR-379/FOXP2 Pathways. Mol Ther Nucleic Acids. 2018;10:331-348.
[18]	Dai W , Yang H , Xu B ,et al.Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) alleviate excessive autophagy of ovarian granular cells through VEGFA/PI3K/AKT/mTOR pathway in premature ovarian failure rat model[J].Journal of Ovarian Research, 2023, 16(1).
[19]	Duan ZN, Dong CG, Liu JH. Circ-ABCB10 promotes growth and metastasis of nasopharyngeal carcinoma by upregulating ROCK1. Eur Rev Med Pharmacol Sci. 2020;24(23):12208-12215.
[20]	Li Y, Gao X, Yang C, Yan H, Li C. CircRNA hsa_circ_0018289 exerts an oncogenic role in cervical cancer progression through miR-1294/ICMT axis. J Clin Lab Anal. 2022;36(5):e24348.
[21]	Deng N, Li L, Gao J, et al. Hsa_circ_0009910 promotes carcinogenesis by promoting the expression of miR-449a target IL6R in osteosarcoma. Biochem Biophys Res Commun. 2018;495(1):189-196.
[22]	Xiao L , Ma X X , Luo J ,et al.Circular RNA circHIPK3 promotes homeostasis of the intestinal epithelium by reducing miR-29b function[J].Gastroenterology, 2021, 161(Suppl 1).
[23]	Yin H , Wang H , Wang M ,et al.CircTCF25 serves as a sponge for miR-206 to support proliferation, migration, and invasion of glioma via the Jak2/p-Stat3/CypB axis[J].Molecular carcinogenesis, 2022, 61(6):558-571.
[24]	He Z, Ruan X, Liu X, et al. FUS/circ_002136/miR-138-5p/SOX13 feedback loop regulates angiogenesis in Glioma. J Exp Clin Cancer Res. 2019;38(1):65.
[25]	Zhao X, Liu Y, Li Z, et al. Linc00511 acts as a competing endogenous RNA to regulate VEGFA expression through sponging hsa-miR-29b-3p in pancreatic ductal adenocarcinoma. J Cell Mol Med. 2018;22(1):655-667.
[26]	Ren L, Ma D, Liu B, et al. Preparation of three-dimensional vascularized MSC cell sheet constructs for tissue regeneration. Biomed Res Int. 2014;2014:301279.
[27]	Yuan Z , Bian Y , Ma X ,et al.LncRNA H19 knockdown in human amniotic mesenchymal stem cells suppresses angiogenesis by associating with EZH2 and activating VASH1[J].Stem Cells and Development, 2019, 28(12).
[28]	Edwards SS, Zavala G, Prieto CP, et al. Functional analysis reveals angiogenic potential of human mesenchymal stem cells from Wharton's jelly in dermal regeneration. Angiogenesis. 2014;17(4):851-866.
[29]	Miao J, Ren Z, Zhong Z, et al. Mesenchymal Stem Cells: Potential Therapeutic Prospect of Paracrine Pathways in Neonatal Infection. J Interferon Cytokine Res. 2021;41(10):365-374.
[30]	Pan L, Lian W, Zhang X, et al. Human circular RNA 0054633 regulates high glucose induced vascular endothelial cell dysfunction through the microRNA 218/roundabout 1 and microRNA 218/heme oxygenase 1 axes. Int J Mol Med. 2018;42(1):597-606.
[31]	Zhang L , Yu D .Exosomes in cancer development, metastasis, and immunity[J].Biochimica et Biophysica Acta (BBA) - Reviews on Cancer, 2019.
[32]	Soltész B, Buglyó G, Németh N, et al. The Role of Exosomes in Cancer Progression. Int J Mol Sci. 2021;23(1):8.
[33]	Li J, Li Z, Jiang P, et al. Circular RNA IARS (circ-IARS) secreted by pancreatic cancer cells and located within exosomes regulates endothelial monolayer permeability to promote tumor metastasis. J Exp Clin Cancer Res. 2018;37(1):177.
[34]	Liang G, Liu Z, Tan L, Su AN, Jiang WG, Gong C. HIF1α-associated circDENND4C Promotes Proliferation of Breast Cancer Cells in Hypoxic Environment. Anticancer Res. 2017;37(8):4337-4343.
[35]	Meng J, Chen S, Han JX, et al. Twist1 Regulates Vimentin through Cul2 Circular RNA to Promote EMT in Hepatocellular Carcinoma. Cancer Res. 2018;78(15):4150-4162.