The Expression And Function Of FOXC1and LncRNA FOXCUT In Oral Squamous Cell Carcinoma

Objective：To preliminarily study the expression of FOXC1（Forkhead box C1，FOXC1）in OSCC（Oral squamous cell carcinoma， OSCC）tissues and Its functionsin Tca8113and SCC-9cells. To analyse correlation between FOXC1and its adjacentlong non coding RNA（long non code RNA，LncRNA）that we named FOXC1promoter upstream transcript（FOXCUT）. To preliminarily study the expression ofFOXCUT in OSCC tissues and Its functions in Tca8113and SCC-9cells. Topreliminarily study the tumor growth inhibition in nude mice which injected Tca8113and SCC-9cells transfected by FOXC1and FOXCUT siRNAs（small interferingRNA, siRNA）.Methods： Employing immunohistochemical techniques（Immunohistochemistry，IHC），Western blot（Western blot，WB）and polymerasechain reaction （Real time Polymerase Chain Reaction， Real time PCR）to detect theFOXC1expression levels in OSCC tissues and corresponding adjacent normal tissues;To detect the expression of the FOXCUT near FOXC1by PCR in the OSCC tissues;And to study the correlation between FOXC1and FOXCUT by Pearson’s correlationcoefficients; To establish siRNA（ssmall interfering RNA, siRNA）by knocking downthe expression of FOXC1and FOXCUT. The valid siRNA was detected by Real timePCR. To research the functions of FOXC1and LncRNA FOXCUT in Tca8113andSCC-9cells transfected with the valid siRNA by using theMethylThiazolylTetrazolium（MTS）Cell Proliferation Assay， Colony FormationAssay， Scratch Wound Healing Assay， CFS formation assay. To verify the FOXC1and FOXCUT functions by constructing BALB/c nude mice model bearing tumorwhich FOXC1siRNA and FOXCUT siRNA inhibit the proliferation of Tca8113cells.Results： Part Ⅰ：1、The expression of FOXC1in OSCC tumor tissues washigher than that of in adjacent normal tissues;2、The downregulation of FOXC1 expression can inhibit the proliferation of oral squamous cell carcinoma Tca8113andSCC-9cell lines;3、The downregulation of FOXC1expression can inhibit theinvasion of oral squamous cell carcinoma Tca8113and SCC-9cell lines;4、Thedownregulation of FOXC1gene expression can reduce the OSCC stem cell function.Part Ⅱ：1、The expression of LncRNA in OSCC tumor tissues was higher thanthat of adjacent normal tissues;2、The downregulation of LncRNA expression caninhibit the proliferation of oral squamous cell carcinoma Tca8113and SCC-9celllines;3、The downregulation of LncRNA expression can inhibit the invasion of oralsquamous cell carcinoma Tca8113and SCC-9cell lines;4、The downregulation ofLncRNA FOXCUT expression can reduce the OSCC stem cell function.Part Ⅲ：1、FOXC1and LncRNA was correlated in gene sequences position;2、The expression FOXC1and LncRNA in OSCC tumor tissues was correlated;3、Thedownregulation of FOXC1expression can not inhibit the FOXCUT;3、 Thedownregulation of LncRNA expression can inhibit the FOXC1;4、FOXC1-LncRNAFOXCUT functioned in the mechanism of OSCC as a gene pair；5、Knockdown ofFOXC1and FOXCUT inhibit the expression of VEGF-A，MMP2，MMP9andMMP7.Part Ⅳ：1、The downregulation of FOXC1and LncRNA FOXCUT expressioncan inhibit the growth of tumors in BALB/c nude mice.2、The mRNA and protinexpression of FOXC1and FOXCUT were down regulated.Conclusion：1、The expression of FOXC1and FOXCUT increased in OSCCtumor tissue，and FOXCUT correlated with FOXC1positively;2、FOXC1andFOXCUT promote the proliferation of tumors and the occurrence and development ofmechanism in OSCC;3、FOXC1-LncRNA gene pair is a kind of new control mode inoccurrence of mechanism in OSCC;4、LncRNA can inhibit the proliferation andinvasion of OSCC cells by regulating FOXC1;5、 The downregulation ofFOXC1-LncRNA expression can inhibit BALB/c tumor growth.