Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China.
Oral Dis. 2022 Apr;28(3):711-722. doi: 10.1111/odi.13771. Epub 2021 Feb 3.
Although integrins have been shown to be associated with proliferation and differentiation in some stem cells, the regulatory effect of integrin α6 (ITGα6) on the human dental pulp stem cells (hDPSCs) has not been reported. Here, we detected the roles of ITGα6 in hDPSCs.
Attached to Cytodex 3 microcarriers, hDPSCs grown under stimulated microgravity (SMG) or conventional culture conditions were measured the proliferation and different gene expression. Further, ITGα6 was silenced in hDPSCs, and its effect on proliferation, differentiation, and cytoskeletal organization was analyzed.
SMG conditions increased the number of Ki67-positive hDPSCs and progression into S phase of cell cycle. WB analysis showed the expression of ITGα6 was upregulated in hDPSCs under SMG conditions. Knockdown of ITGα6 decreased the expression of stemness markers, CD105 and STRO-1 in hDPSCs, but promoted the osteogenic and odontogenic differentiation by increased ALP expression and Alizarin Red nodules. Moreover, RNA-seq demonstrated that RHO/ROCK signaling pathway upregulated silencing ITGα6-hDPSCs. Treatment with Y-27632 inhibited the effect of ITGα6 depletion on hDPSCs stemness, rearranged the cytoskeleton, promoted the pluripotency, proliferation ability, and inhibited the differentiation.
ITGα6 promotes hDPSCs stemness via inhibiting RHO/ROCK and restoring cytoskeleton.
尽管整合素已被证明与某些干细胞的增殖和分化有关，但整合素 α6（ITGα6）对人牙髓干细胞（hDPSCs）的调节作用尚未报道。在这里，我们检测了 ITGα6 在 hDPSCs 中的作用。
附着在 Cytodex 3 微载体上，在受刺激的微重力（SMG）或常规培养条件下生长的 hDPSCs 被测量增殖和不同基因表达。进一步，沉默 hDPSCs 中的 ITGα6，并分析其对增殖、分化和细胞骨架组织的影响。
SMG 条件增加了 Ki67 阳性 hDPSCs 的数量，并促进了细胞周期的 S 期进程。WB 分析显示，SMG 条件下 hDPSCs 中 ITGα6 的表达上调。ITGα6 的敲低降低了 hDPSCs 中干细胞标志物 CD105 和 STRO-1 的表达，但通过增加 ALP 表达和茜素红结节促进成骨和成牙分化。此外，RNA-seq 表明 RHO/ROCK 信号通路上调了沉默 ITGα6-hDPSCs。用 Y-27632 处理抑制了 ITGα6 耗竭对 hDPSCs 干性的影响，重排细胞骨架，促进多能性、增殖能力，并抑制分化。
ITGα6 通过抑制 RHO/ROCK 和恢复细胞骨架来促进 hDPSCs 的干性。
