Expression And Significance Of OPG,RANKL And RANK Proteins In Osteosarcoma Of The Jaw

Objective:To investigate the expression of osteoprotegerin(OPG),receptor activator of NF-?B(RANK)and its only ligand receptor activator of NF-?B Ligand(RANKL)in osteosarcoma of the jaw,and to analyse the role of OPG/RANKL/RANK signaling axis in the occurrence and development of osteosarcoma of the jaw and its association with metastasis of osteosarcoma,with the purpose of providing reference for the clinic therapy of osteosarcoma of the jaw.Methods:(1)Case collection:32 cases of osteosarcoma of the jaw paraffin-embedded specimens(11 cases with metastasis and 21 cases without metastasis)and 10 cases of the odontogenic keratocyst paraffin-embedded specimens were seclected and confirmed by pathology(Odontogenic keratocyst can cause obvious damageto the bone of the jaw,presenting the biological behavior of infiltrating growth,and have significant recurrence and canceration ability.Therefore,compared with the general jaw cyst,the odontogenic keratocyst have certain particularity,so they were selected as the control group).(2)Experimental grouping:routine pathological sectioning were performed on the included osteosarcoma of the jaw and keratocystic odontogenic tumor tissues,and the cases were divided into?osteosarcoma of the jaw with metastasis group;?osteosarcoma of the jaw without metastasis group;?odontogenic keratocyst group.(3)The morphology and distribution of osteoclasts in osteosarcoma of the jaw were observed by TRAP immunohistochemistry.(4)Immunohistochemistry(IHC)was used to detect the expression of OPG,RANKL and RANK in osteosarcoma of the jaw with metastasis group,osteosarcoma of the jaw without metastasis group and odontogenic keratocyst group respectively.(5)Interpretation of results:5 fields were selectedrandomly from each slice.The immunohistochemical results of this experiment were interpreted by Immunoreactive Score(IRS)method,that is to say Staining Intensity(SI):No staining is 0,primrose yellow is 1 point,light brown is 2 points,dark brown is 3 points;Percentage of Positive Cells(PP):Negative is 0,1%-25%is 1 point,26%-50%is 2 points,51%-75%is 3 points,>75%is 4 points;IRS=SI*PP,IRS=0 means Negative(-);IRS=1-4 means Low positive(+);IRS=5-8 means Positive(++);IRS=9-12 means High positive(+++).(6)Deta processing:Statistical analysis was performed on the data using SPSS 23.0 software.The ranked data was analyzed by multiple independent samples of the Kruskal-Wallis rank sum test,Spearman non-parametric correlation analysis was used to analyze the level correlation,take the level of hypothesis test ?=0.05,the difference was statistically significant at P<0.05.Results:(1)TRAP staining showed that there were a large number of osteoclasts in the osteosarcoma of the jaw tissue,the staining positive area expressed as aubergine,with dense distribution of osteoclasts,polygonal morphology,various sizes and multiple nuclei in a cell.(2)The results of OPG immunohistochemical staining showed that OPG protein was mainly located in the extracellular matrix.There were differences in the expression intensity of OPG in these three groups(P<0.05).There was no statiscally significant difference in the expression intensity of OPG between osteosarcoma of the jaw with metastasis group and odontogenic keratocyst group(P>0.05).The expression intensity of OPG in the osteosarcoma of the jaw without metastasis group was higher than that in odontogenic keratocyst group,and the difference was statistically significant(P<0.05).The expression intensity of OPG in the osteosarcoma of the jaw without metastasis group was higher than that inthe osteosarcoma of the jaw with metastasis group,and the difference was statistically significant(P<0.05).(3)The results of RANKL immunohistochemical staining showed that RANKL protein was mainly located in cytoplasm and cell membrane.There were differences in the expression intensity of RANKL in these three groups(P<0.05).The expression intensity of RANKL in the osteosarcoma of the jaw with metastasis group was higher than that in odontogenic keratocyst group,and the difference was statistically significant(P<0.05).The expression intensity of RANKL in the osteosarcoma of the jaw without metastasis group was higher than that in odontogenic keratocyst group,and the difference was statistically significant(P<0.05).The expression intensity of RANKL in the osteosarcoma of the jaw with metastasis group was higher than that inthe osteosarcoma of the jaw without metastasis group,and the difference was statistically significant(P<0.05).(4)The results of RANK immunohistochemical staining showed that RANK protein was mainly located in cytoplasm and cell membrane.There were differences in the expression intensity of RANK in these three groups(P<0.05).The expression intensity of RANK in the osteosarcoma of the jaw with metastasis group was higher than that in odontogenic keratocyst group,and the difference was statistically significant(P<0.05).The expression intensity of RANK in the osteosarcoma of the jaw without metastasis group was higher than that in odontogenic keratocyst group,and the difference was statistically significant(P<0.05).The expression intensity of RANK in the osteosarcoma of the jaw with metastasis group was higher than that inthe osteosarcoma of the jaw without metastasis group,and the difference was statistically significant(P<0.05).(5)Correlation analysis of the expression intensity of OPG,RANKL and RANK in the osteosarcoma of the jaw with metastasis group:there was no correlation between OPG and RANKL(P=0.053>0.05);no correlation between OPG and RANK(P=0.079>0.05);a significantly positive correlation between OPG and RANKL(rs=0.980,P<0.001)(6)Correlation analysis of the expression intensity of OPG,RANKL and RANK in the osteosarcoma of the jaw without metastasis group:there was a significantly negative correlation between OPG and RANKL(rs=-0.771,P<0.001);a significantly negative correlation between OPG and RANK(rs=-0.841,P<0.001);a significantly positive correlation between OPG and RANKL(rs=0.862,P<0.001).(7)Correlation analysis of the expression intensity of OPG,RANKL and RANK in the odontogenic keratocyst group:there was a significantly negative correlation between OPG and RANKL(rs=-0.75,P=0.012<0.05);no correlation between OPG and RANK(P=0.052>0.05);a significantly positive correlation between OPG and RANKL(rs=0.082,P=0.001<0.05).Conclusion:(1)Osteoclasts are abundant and actively differentiated in the osteosarcoma of the jaw,it may be closely related the resorption and destruction of the jaw.(2)The expression of OPG,RANKL and RANK can be dectected in the tissues of osteosarcoma of the jaw,and the expression levels of RANKL and RANK were higher than that of odontogenic keratocyst tissue.(3)The expression levels of RANKL and RANK were significantly enhanced during the metastasis of osteosarcoma of the jaw.(4)There was a positive correlation between the expression levels of RANKL and RANK in the osteosarcoma of the jaw tissue.(5)OPG/RANKL/RANK signal axis is involved in the occurrence,development and metastasis of osteosarcoma of the jaw.