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Whether these mutations affect osteoblast differentiation remains to be determined. This study aimed to investigate the effects of <jats:italic>WNT1<\/jats:italic> c.110\u2009T&gt;C and c.505G&gt;T mutations on osteoblast function, gene expression, and pathways involved in OI.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Empty vector (negative control), wild-type <jats:italic>WNT1<\/jats:italic>, <jats:italic>WNT1<\/jats:italic> c.110\u2009T&gt;C, <jats:italic>WNT1<\/jats:italic> c.505G&gt;T, and <jats:italic>WNT1<\/jats:italic> c.884C&gt;A (positive control) mutant plasmids were constructed and transfected into preosteoblast (MC3T3-E1) cells to investigate their effect on osteoblast differentiation. The expressions of osteoblast markers, including <jats:italic>BMP2<\/jats:italic>, <jats:italic>RANKL<\/jats:italic>, osteocalcin, and alkaline phosphatase (ALP), were determined using quantitative real-time polymerase chain reaction (RT-qPCR), western blotting (WB), enzyme-linked immunosorbent assay, and ALP staining assay, respectively. The mRNA and protein expression levels of <jats:italic>WNT1<\/jats:italic> or the expression levels of the relevant proteins involved in the WNT1\/\u03b2-catenin signaling pathway were also determined using RT-qPCR, WB, and immunofluorescence (IF) assays after the different plasmids were transfected into MC3T3-E1 cells.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Compared with those in the wild-type group, in the mutation groups, the mRNA and protein expression levels of <jats:italic>BMP2<\/jats:italic> were suppressed, the expressions of osteocalcin and ALP were inhibited, and the mRNA and protein expression levels of <jats:italic>RANKL<\/jats:italic> were enhanced in MC3T3-E1 cells. WB and IF assays revealed that the protein expression levels of <jats:italic>WNT1<\/jats:italic> in MC3T3-E1 cells were downregulated in the mutation groups compared with those in the wild-type <jats:italic>WNT1<\/jats:italic> group. Furthermore, the expression levels of nonphosphorylated \u03b2-catenin (non-p-\u03b2-catenin) and phosphorylated GSK-3\u03b2 (p-GSK-3\u03b2) were downregulated in the mutation groups compared with those in the wild-type group. However, no significant changes in the expression level of non-p-\u03b2-catenin or p-GSK-3\u03b2 were observed in the mutation groups.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p><jats:italic>WNT1<\/jats:italic> c.110\u2009T&gt;C and c.505G&gt;T mutations may alter the proliferation and osteogenic phenotype of MC3T3-E1 linked to the progression of OI via the inhibition of the WNT1\/\u03b2-catenin signaling pathway. This is the first study to confirm the effect of <jats:italic>WNT1<\/jats:italic> c.110\u2009T&gt;C and c.505G&gt;T missense mutations on osteoblast differentiation and propose a new molecular mechanism for OI development.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s13018-021-02495-2","type":"journal-article","created":{"date-parts":[[2021,6,2]],"date-time":"2021-06-02T07:03:56Z","timestamp":1622617436000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Effects of WNT1 c.110\u2009T&gt;C and c.505G&gt;T mutations on osteoblast differentiation via the WNT1\/\u03b2-catenin signaling pathway"],"prefix":"10.1186","volume":"16","author":[{"given":"Bashan","family":"Zhang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rong","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenfeng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xueming","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beijing","family":"Luo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zinian","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xibo","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aijiao","family":"Ding","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,6,2]]},"reference":[{"issue":"8","key":"2495_CR1","doi-asserted-by":"publisher","first-page":"1042","DOI":"10.1038\/ejhg.2015.81","volume":"23","author":"K Lindahl","year":"2015","unstructured":"Lindahl K, Astrom E, Rubin CJ, Grigelioniene G, Malmgren B, Ljunggren O, et al. 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