Effect of LOC103693069 on hypoxic apoptosis of bone marrow mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

WANG Chuan , ZHANG Fei ,  PENG Wuxun , WANG Tao , XIE Zhihong , YAN Yanglin , QIU Hanke

Emergency Department, the Affiliated Hospital of Guizhou Medical University, Guiyang Guizhou, 550004, P. R. China;

Corresponding author：

PENG Wuxun, Email: wangchuan22222@163.com

Keywords：

Bone marrow mesenchymal stem cells; long non-coding RNA; hypoxic apoptosis; rat

DOI：

10.7507/1002-1892.202103092

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the effect of LOC103693069 on hypoxic apoptosis of bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs from 1-week-old Sprague Dawley rat bone marrow were isolated, cultured, and passaged by the whole bone marrow adherent culture method. After identification of adipogenic, chondrogenic, and osteogenic differentiation, the 3rd generation cells were treated with hypoxia under 5%O₂, 1%O₂, and anaerobic conditions. After 48 hours, the cell viability, apoptosis, and apoptosis-related proteins [hypoxia inducible factor 1α (HIF-1α), Caspase-3, B cell lymphoma/leukemia 2 (Bcl-2)] expressions were detected, and normal BMSCs were used as controls. Based on the research results, the concentration group with the most obvious apoptosis was selected and used for subsequent experiments. After 48 hours of hypoxia treatment, BMSCs were taken and analyzed by gene chip and real-time fluorescence quantitative PCR (qRT-PCR) to screen the most significantly down-regulated gene and construct their high-expression, low-expression, and negative control lentiviruses; BMSCs were transfected with the different lentiviruses, respectively. After qRT-PCR detection confirmed that the transfection was successful, the BMSCs were treated with hypoxia for 48 hours to observe the cell viability and the expressions of apoptosis-related proteins. Results After cell viability, apoptosis, and apoptosis-related proteins were detected, cell apoptosis was the most significant under anaerobic conditions after 48 hours. The above indicators were significantly different from other groups (P<0.05), and this group was used for treatment conditions for subsequent experiments. Gene chip analysis showed that after 48 hours of hypoxia treatment, AC125847.1, LOC102547753, AABR07017208.2, and LOC103693069 were significantly down-regulated in BMSCs, and the expressions of LOC103693069 was the most significant down-regulation detected by qRT-PCR (P<0.05). It was selected to construct lentivirus and transfect BMSCs. Afterwards, qRT-PCR detection showed the successful transfection into the cells. After hypoxia treatment, the apoptosis rate and the expressions of apoptosis-related proteins of BMSCs overexpressed by the gene were significantly reduced (P<0.05). Conclusion LOC103693069 can relieve the hypoxic apoptosis of BMSCs.

Citation： WANG Chuan, ZHANG Fei, PENG Wuxun, WANG Tao, XIE Zhihong, YAN Yanglin, QIU Hanke. Effect of LOC103693069 on hypoxic apoptosis of bone marrow mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2022, 36(3): 362-369. doi: 10.7507/1002-1892.202103092 Copy

1.	Zhang J, Feng Z, Wei J, et al. Repair of critical-sized mandible defects in aged rat using hypoxia preconditioned BMSCs with up-regulation of Hif-1α. Int J Biol Sci, 2018, 14(4): 449-460.
2.	Chai M, Gu C, Shen Q, et al. Hypoxia alleviates dexamethasone-induced inhibition of angiogenesis in cocultures of HUVECs and rBMSCs via HIF-1α. Stem Cell Res Ther, 2020, 11(1): 343. doi: 10.1186/s13287-020-01853-x.
3.	Li YM, Chen GH, Yu DS, et al. Ebselen rescues high glucose suppressed osteogenic differentiation of BMSCs by the activation of PI3K AKT pathway. Clinical Oral Implants Research, 2010, 30(S19): 54. doi: 10.1111/clr.16_13509.
4.	Zhang M, Shi X, Wu J, et al. CoCl₂ induced hypoxia enhances osteogenesis of rat bone marrow mesenchymal stem cells through cannabinoid receptor 2. Arch Oral Biol, 2019, 108: 104525. doi: 10.1016/j.archoralbio.2019.104525.
5.	Zhu J, Fu Q, Shao J, et al. Over-expression of MEG3 promotes differentiation of bone marrow mesenchymal stem cells into chondrocytes by regulating miR-129-5p/RUNX1 axis. Cell Cycle, 2021, 20(1): 96-111.
6.	Yuan S, Zhang C, Zhu Y, et al. Neohesperidin ameliorates steroid-induced osteonecrosis of the femoral head by inhibiting the histone modification of lncRNA HOTAIR. Drug Des Devel Ther, 2020, 14: 5419-5430.
7.	Wang W, Wang Y, Deng G, et al. Transplantation of hypoxic-preconditioned bone mesenchymal stem cells retards intervertebral disc degeneration via enhancing implanted cell survival and migration in rats. Stem Cells Int, 2018, 2018: 7564159. doi: 10.1155/2018/7564159.
8.	Bernard O, Jeny F, Uzunhan Y, et al. Mesenchymal stem cells reduce hypoxia-induced apoptosis in alveolar epithelial cells by modulating HIF and ROS hypoxic signaling. Am J Physiol Lung Cell Mol Physiol, 2018, 314(3): L360-L371.
9.	Chen L, Yang Y, Peng X, et al. Transcription factor YY1 inhibits the expression of THY1 to promote interstitial pulmonary fibrosis by activating the HSF1/miR-214 axis. Aging (Albany NY), 2020, 12(9): 8339-8351.
10.	Wu K, Zou J, Lin C, et al. MicroRNA-140-5p inhibits cell proliferation, migration and promotes cell apoptosis in gastric cancer through the negative regulation of THY1-mediated Notch signaling. Biosci Rep, 2019, 39(7): BSR20181434. doi: 10.1042/BSR20181434.
11.	Ishifune C, Maekawa Y, Nishida J, et al. Notch signaling regulates the development of a novel type of Thy1-expressing dendritic cell in the thymus. Eur J Immunol, 2011, 41(5): 1309-1320.
12.	Mo W, Li Y, Chang W, et al. The role of LncRNA H19 in MAPK signaling pathway implicated in the progression of bronchopulmonary dysplasia. Cell Transplant, 2020, 29: 963689720918294. doi: 10.1177/0963689720918294.
13.	Guan CJ, Wang Y. LncRNA CASC9 attenuates lactate dehydrogenase-mediated oxidative stress and inflammation in spinal cord injury via sponging miR-383-5p. Inflammation, 2021, 44: 923-933.
14.	Ahmad I, Valverde A, Naqvi RA, et al. Long non-coding RNAs RN7SK and GAS5 regulate macrophage polarization and innate immune responses. Front Immunol, 2020, 11: 604981. doi: 10.3389/fimmu.2020.604981.
15.	Yao XY, Liu JF, Luo Y, et al. LncRNA HOTTIP facilitates cell proliferation, invasion, and migration in osteosarcoma by interaction with PTBP1 to promote KHSRP level. Cell Cycle, 2021, 20(3): 283-297.

1. Zhang J, Feng Z, Wei J, et al. Repair of critical-sized mandible defects in aged rat using hypoxia preconditioned BMSCs with up-regulation of Hif-1α. Int J Biol Sci, 2018, 14(4): 449-460.
2. Chai M, Gu C, Shen Q, et al. Hypoxia alleviates dexamethasone-induced inhibition of angiogenesis in cocultures of HUVECs and rBMSCs via HIF-1α. Stem Cell Res Ther, 2020, 11(1): 343. doi: 10.1186/s13287-020-01853-x.
3. Li YM, Chen GH, Yu DS, et al. Ebselen rescues high glucose suppressed osteogenic differentiation of BMSCs by the activation of PI3K AKT pathway. Clinical Oral Implants Research, 2010, 30(S19): 54. doi: 10.1111/clr.16_13509.
4. Zhang M, Shi X, Wu J, et al. CoCl₂ induced hypoxia enhances osteogenesis of rat bone marrow mesenchymal stem cells through cannabinoid receptor 2. Arch Oral Biol, 2019, 108: 104525. doi: 10.1016/j.archoralbio.2019.104525.
5. Zhu J, Fu Q, Shao J, et al. Over-expression of MEG3 promotes differentiation of bone marrow mesenchymal stem cells into chondrocytes by regulating miR-129-5p/RUNX1 axis. Cell Cycle, 2021, 20(1): 96-111.
6. Yuan S, Zhang C, Zhu Y, et al. Neohesperidin ameliorates steroid-induced osteonecrosis of the femoral head by inhibiting the histone modification of lncRNA HOTAIR. Drug Des Devel Ther, 2020, 14: 5419-5430.
7. Wang W, Wang Y, Deng G, et al. Transplantation of hypoxic-preconditioned bone mesenchymal stem cells retards intervertebral disc degeneration via enhancing implanted cell survival and migration in rats. Stem Cells Int, 2018, 2018: 7564159. doi: 10.1155/2018/7564159.
8. Bernard O, Jeny F, Uzunhan Y, et al. Mesenchymal stem cells reduce hypoxia-induced apoptosis in alveolar epithelial cells by modulating HIF and ROS hypoxic signaling. Am J Physiol Lung Cell Mol Physiol, 2018, 314(3): L360-L371.
9. Chen L, Yang Y, Peng X, et al. Transcription factor YY1 inhibits the expression of THY1 to promote interstitial pulmonary fibrosis by activating the HSF1/miR-214 axis. Aging (Albany NY), 2020, 12(9): 8339-8351.
10. Wu K, Zou J, Lin C, et al. MicroRNA-140-5p inhibits cell proliferation, migration and promotes cell apoptosis in gastric cancer through the negative regulation of THY1-mediated Notch signaling. Biosci Rep, 2019, 39(7): BSR20181434. doi: 10.1042/BSR20181434.
11. Ishifune C, Maekawa Y, Nishida J, et al. Notch signaling regulates the development of a novel type of Thy1-expressing dendritic cell in the thymus. Eur J Immunol, 2011, 41(5): 1309-1320.
12. Mo W, Li Y, Chang W, et al. The role of LncRNA H19 in MAPK signaling pathway implicated in the progression of bronchopulmonary dysplasia. Cell Transplant, 2020, 29: 963689720918294. doi: 10.1177/0963689720918294.
13. Guan CJ, Wang Y. LncRNA CASC9 attenuates lactate dehydrogenase-mediated oxidative stress and inflammation in spinal cord injury via sponging miR-383-5p. Inflammation, 2021, 44: 923-933.
14. Ahmad I, Valverde A, Naqvi RA, et al. Long non-coding RNAs RN7SK and GAS5 regulate macrophage polarization and innate immune responses. Front Immunol, 2020, 11: 604981. doi: 10.3389/fimmu.2020.604981.
15. Yao XY, Liu JF, Luo Y, et al. LncRNA HOTTIP facilitates cell proliferation, invasion, and migration in osteosarcoma by interaction with PTBP1 to promote KHSRP level. Cell Cycle, 2021, 20(3): 283-297.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of LOC103693069 on hypoxic apoptosis of bone marrow mesenchymal stem cells

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