南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (9): 1394-1399.doi: 10.12122/j.issn.1673-4254.2021.09.15

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RUNX2/LAPTM5在小鼠颅骨前成骨细胞矿化诱导中的表达

邢 磊，耿远明，李文昊，林丽佳，徐平平   

  1. 广州医科大学附属口腔医院种植科//广州市口腔再生医学基础与应用研究重点实验室，广东 广州 510182；南方医科大学珠江医院口腔科，广东 广州 510282；南方医科大学口腔医院，广东 广州 510280
  • 出版日期:2021-09-20 发布日期:2021-09-30

Expression of RUNX2 /LAPTM5 inMC3T3-E1osteoblastic cellswith inducedmineralization

XING Lei, GENG Yuanming, LI Wenhao, LIN Lijia, XU Pingping   

  1. Department of Oral Implantology, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangzhou Key Laboratory of Basic and Applied Research in Oral Regenerative Medicine, Guangzhou 510182, China; Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China; Stomatological Hospital, Southern Medical University, Guangzhou 510280, China
  • Online:2021-09-20 Published:2021-09-30

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摘要： 目的 探讨RUNX2/LAPTM5在矿化诱导过程中的表达与成骨及溶酶体的相关性。方法 矿化诱导MC3T3-E1，对照组不做处理，茜素红染色检测矿化情况，碱性磷酸酶染色检测成骨分化情况。RT-qPCR及Western blot检测分化0-5 d RUNX2及LAPTM5的基因及蛋白表达。过表达与干扰RUNX2/LAPTM5的表达后，Western blot检测RUNX2、LAPTM5的表达。过表达与干扰LAPTM5的表达后，Western blot检测成骨相关基因碱性磷酸酶、骨钙素 的表达。结果 倒置显微镜下观察，茜素红染色矿化结节计数随时间变化逐渐增多，矿化结节的大小也逐渐变大；碱性磷酸酶染色蓝紫色颗粒计数随时间逐渐增加。RT-qPCR及Western blot结果显示RUNX2及LAPTM5的表达，其在成骨矿化过程中呈上升趋势（P<0.001）。过表达与干扰RUNX2影响LAPTM5表达（P<0.05）；过表达与干扰LAPTM5对RUNX2的影响不显著。过表达与干扰LAPTM5影响了成骨的表达（P< 0.01）。结论 RUNX2/LAPTM5可能参与了成骨细胞分化调节，RUNX2可能参与LAPTM5的表达调控。RUNX2/LAPTM5可能在溶酶体参与成骨矿化的过程中起到桥梁作用。

关键词: RUNX2；LAPTM5；成骨；溶酶体

Abstract: Objective To investigate the association of the expressions of RUNX2/LAPTM5 with osteogenesis and lysosomes in osteoblastic cells during mineralization induction. Methods MC3T3- E1 cells cultured in osteogenic induction medium was examined for mineralization and osteogenic differentiation using Alizarin red staining and alkaline phosphatase (ALP) staining, respectively. RT- qPCR and Western blotting were used to detect the mRNA and protein expressions of Runx2 and LAPTM5 in the cells during osteogenic induction for 5 days. The effects of overexpression and interference of RUNX2/LAPTM5 on the expressions of ALP and osteocalcin (OCN) in the cells were examined with Western blotting. Results MC3T3- E1 cells cultured in osteogenic induction medium showed an increased number of mineralized nodules over time, and the size of the mineralized nodules increased as the culture time extended; the number of purple-blue granules stained by ALP also increased gradually with time. RT- qPCR and Western blotting showed that the expressions of RUNX2 and LAPTM5 in the cells increased progressively during osteogenic mineralization (P<0.001). Overexpression and interference of RUNX2 obviously affected LAPTM5 expression in the cells (P<0.05); modulation of LAPTM5 expression did not significantly affect RUNX2 expression but caused significant changes in ALP and OCN expressions (P<0.01). Conclusion RUNX2 /LAPTM5 may participate in the regulation of osteoblast differentiation, and RUNX2 may be involved in the regulation of LAPTM5 expression. RUNX2 /LAPTM5 may play a mediating role in the process of osteogenic mineralization involving lysosomes.

Key words: RUNX2; LAPTM5; osteogenesis; lysosomes