| Head and neck malignancy(HNC)is one of the most common human cancers,and oral squamous cell carcinoma(OSCC)is the most common type.More than 400,000 people are affected every year.The 5-year survival rate of patients does not exceed 50%.In this study,based on the targeted gene editing of heat shock protein 47(HSP47)gene,a model of HSP47 overexpression and knockout of SCC9 and SAS tongue cancer cells was constructed,and the model group’s proliferation,invasion,migration,and migration compared with wild-type tongue cancer cells were studied.Changes in apoptotic function.Immunofluorescence technique was used to study the effect of HSP47 on collagen and endoplasmic reticulum stress protein.And based on HSP47,the enrichment analysis of GSEA signaling pathway was carried out,and the relationship between HSP47 and PI3K/AKT,ERK/MAPK,TGF-β,ECM related pathways was discussed.Objectives:1.To study the effect of HSP47 on the proliferation,invasion,migration and apoptosis of tongue cancer cells;2.To study the effect of HSP47 on the maturation and secretion of collagen in tongue cancer cells and the effect on endoplasmic reticulum stress;3.To study the effect of HSP47 on the apoptosis mechanism of tongue cancer cells;4.To study whether HSP47 promotes the progression of tongue cancer by mediating PI3K/AKT,ERK/MAPK,TGF-β,and ECM pathways.、Methods:1.The results of qPCR experiments showed that the expression of HSP47 relative m RNA in OSCC patients’ cancer tissues was higher than that in adjacent tissues,and the difference was statistically significant(P<0.05);2.CCK-8 experiment and clone formation experiment showed that the proliferation ability of the OE group was significantly greater than that of the wild-type WT group,and the difference was statistically significant(P<0.05);the proliferation ability of the S1 group was lower than that of the wild-type tongue cancer cell group,and the difference was statistically significant Significance(P<0.05);3.Scratch experiment,transwell migration and invasion experiments showed that the migration and invasion ability of OE group was significantly greater than that of WT group,the difference was statistically significant(P<0.05);the migration and invasion ability of S1 group was lower than that of WT group,and the difference was statistically significant(P<0.05);4.Apoptosis experiment results showed that the apoptotic ability of the OE group was less than that of the WT group,and the difference was statistically significant(P<0.05);the apoptotic ability of the S1 group was greater than that of the WT group,and the difference was statistically significant(P<0.05);5.In SCC9 and SAS cells,the expression of collagen I and II in the OE group was higher than that in the WT group,and the distribution was more diffuse,not only limited to the ER.In the S1 group,the opposite is true;6.In SCC9 and SAS cells,the expression of endoplasmic reticulum stress protein phosphorylation IRE1 and PERK in S1 group was higher than that in WT and OE groups.7.The results of the Western Blot experiment showed that the expression of Bcl-2,Bcl-XL,and Mcl-1 in the S1 group decreased,and the expression of Bax,Bad,and PUAM increased.In the OE group,the expression of Bcl-2,Bcl-XL,and Mcl-1 decreased,and the expression of Bax,Bad,and PIUAM increased;8.HSP47 GSEA analysis results show that HSP47 is related to PI3K/AKT,ERK/MAPK,TGF-β and ECM pathways.Western Blot experiment found that in SCC9 and SAS,the protein expressions of mmp2,mmp9,AKT,p-AKT,ERK1/2,p-MAPK,and TGF-β in the OE group were higher than those in the WT group,and the S1 group was lower than the WT group.Results:1.The results of qPCR experiments showed that the expression of HSP47 relative m RNA in OSCC patients’ cancer tissues was higher than that in adjacent tissues,and the difference was statistically significant(P<0.05);2.CCK-8 experiment and clone formation experiment showed that the proliferation ability of the OE group was significantly greater than that of the wild-type WT group,and the difference was statistically significant(P<0.05);the proliferation ability of the S1 group was lower than that of the wild-type tongue cancer cell group,and the difference was statistically significant Significance(P<0.05);3.Scratch experiment,transwell migration and invasion experiments showed that the migration and invasion ability of OE group was significantly greater than that of WT group,the difference was statistically significant(P<0.05);the migration and invasion ability of S1 group was lower than that of WT group,and the difference was statistically significant(P<0.05);4.Apoptosis experiment results showed that the apoptotic ability of the OE group was less than that of the WT group,and the difference was statistically significant(P<0.05);the apoptotic ability of the S1 group was greater than that of the WT group,and the difference was statistically significant(P<0.05);5.In SCC9 and SAS cells,the expression of collagen I and II in the OE group was higher than that in the WT group,and the distribution was more diffuse,not only limited to the ER.In the S1 group,the opposite is true;6.In SCC9 and SAS cells,the expression of endoplasmic reticulum stress protein phosphorylation IRE1 and PERK in S1 group was higher than that in WT and OE groups.7.The results of Western Blot experiment showed that the expressions of Bcl-2,Bcl-XL,and Mcl-1 decreased in the S1 group,while the expressions of Bax,Bad,and PUAM increased.In the OE group,the expression of Bcl-2,Bcl-XL,and Mcl-1 decreased and the expression of Bax,Bad and PIUAM increased;8.HSP47 GSEA analysis results show that HSP47 is related to PI3K/AKT,ERK/MAPK,TGF-β and ECM pathways.Western Blot experiment found that in SCC9 and SAS,the protein expressions of mmp2,mmp9,AKT,p-AKT,ERK1/2,p-MAPK,and TGF-β in the OE group were higher than those in the WT group,and the S1 group was lower than the WT group.Conclusions:1.HSP47 promotes the proliferation,invasion and migration of tongue cancer cells,and inhibits their apoptosis;2.HSP47 affects the expression of collagen and endoplasmic reticulum stress-related proteins in tongue cancer cells;3.HSP47 promotes the development of tongue cancer by inhibiting the apoptosis of tongue cancer cells;4.HSP47 promotes the progression of tongue cancer through AKT,MAPK,TGF-β and ECM pathways in tongue cancer cells. |
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