| Background and objectiveOral submucous fibrosis(OSF)is a more common clinical oral mucosal disease,with a potential tendency to cancer.Most patients with OSF have the habit of chewing betel nut,which is also one of the main pathogenic factors.OSF mainly occurs in India,Pakistan,China and other Southeast Asian countries and regions,and in China,it is mainly seen in Hunan,Hainan,Guangdong,Guangxi,Taiwan and other central and southern provinces.In recent years,the population mobility has gradually increased,and the incidence of OSF has also been reported in other provinces,which has gradually become a national health problem.Therefore,studying the cause and pathogenesis of its occurrence is the key to the prevention and treatment of OSF.The literature reported that DKK3 is closely associated with the development of many tumors,but the reported conclusions are different,which has been shown to have a tumor suppressive effect in some tumors and shows its promoting effect in others.However,there are few studies on OSF and oral squamous cell carcinoma(OSCC).At present,there are no studies on OSF combined with OSCC,and no studies on DKK3 in arecoline induced OSF cells.Our research group previously conducted a study on the expression of DKK3 in OSF and OSCC tissues,and found that with the aggravation of OSF,the expression of DKK3 decreased,and the expression sites were different,suggesting that DKK3 may be related to the occurrence and development of OSF and carcinogenesis,but the reason and mechanism are still unclear.Based on the important role of Myofibroblasts(MFB)in OSF,arecan can promote the differentiation of fibroblasts into MFB expressing α-smooth muscle actin(α-SMA)phenotype.In this study,arecoline-induced MFB,oral squamous cell carcinoma cells,OSF tissues,OSF with OSCC and OSCC tissues were used as experimental objects to analyze the expression of DKK3 in different cells and tissues and its changes in OSF and carcinogenesis.It provides experimental and theoretical basis for further study on the role and mechanism of DKK3 in the development and carcinogenesis of OSF.MethodsBased on the current market without OSF cells available,MFB were used as the OSF cell model.(1)Arecoline induces the differentiation of oral mucosal fibroblasts(FB)into MFB.Fresh normal oral mucosa(NOM)tissues were collected for FB isolation,primary culture and passage,and FB was induced by apline at the concentration of 0ug/m L,20ug/m L,40ug/ml and 60ug/ m L respectively.q RT-PCR and Western Blot were used to detect the expression of α-SMA in arecoline-induced MFB cells.(2)The tissue and clinicopathological data of 47 OSF patients(16 in early stage,16 in metaphase,and 15 in advanced stage),16 OSF with OSCC,15 OSCC alone,and 15 NOM were collected as experimental specimens.(3)The pression of DKK3 gene and protein in FB,MFB and oral tumor cells(CAL27,SCC9,SCC25)were measured by q RT-PCR and Western Blot,respectively.(4)DKK3 expression was detected in OSF,OSF with OSCC,OSCC alone and NOM samples,and the relationship with clinical and pathological parameters was analyzed.(5)Statistical analysis was performed using IBM SPSS Statistics26.0 and Graph Pad Prism 9 software,and DKK3 expression in experimental and control groups was compared using Chi-square test or Fisher exact probability method.P<0.05 was considered to be statistically significant.Results(1)The expression of α-SMA m RNA and protein in MFB cells induced by 20ug/ m L arecaloid was significantly higher than that in FB cell group,0ug/ m L group,40ug/ m L group and 60ug/ m L group(P<0.05).Therefore,in this study,20ug/ m L arecaloid concentration was used to induce FB differentiation into MFB in oral mucosa.(2)The expression of DKK3 m RNA and protein in MFB cells was significantly lower than that in FB cells of control group,and the expression in OSCC cells was significantly higher than that in FB cells of control group(P<0.05).(3)The positive expression rate of DKK3 was 73.33%(11/15)in NOM,29.79%(14/47)in OSF,75%(12/16)in OSF and 93.33%(14/15)in OSCC.The positive expression rate of DKK3 in OSF was lower than that of NOM,and the difference was statistically significant(P<0.05).There was no significant difference in DKK3 expression in early,middle and late OSF.The positive expression rate of OSF with OSCC and OSCC alone was higher than that of NOM,but the difference was not statistically significant(P>0.05).The positive expression rates in OSF were lower than those in OSF with OSCC and OSCC alone,and the difference was statistically significant(P<0.05).The positive expression rate of OSF with OSCC was lower than that of OSCC,but the difference was not statistically significant(P>0.05).ConclusionBased on the fact that the expression level of DKK3 in MFB was significantly lower than that of FB in cells,and higher than that in oral cancer cells;The expression rate of DKK3 in OSF tissues was also significantly lower than that in NOM group,OSF with OSCC group and OSCC group,suggesting that DKK3 may be a potential marker for predicting the occurrence of OSF,but not as a marker of OSF canceration.There are 12 figures,22 tables and 87 references... |
PDF Full Text Request