| Objective:Neuroblastoma(NB)is a malignant solid tumor originating from the neural crest,high malignancy and poor prognosis.The improvement of the early diagnosis level can improve the survival rate of the children.Besides the N-myc gene,there is still a lack of effective tumor markers.Nestin(nestin)is a marker of neural stem cells.Studies have shown that nestin is expressed in many tumors and tumor stem cells.In this study,we detected the expression of nestin in 42 cases of neuroblastoma in children,and further understood the correlation between nestin expression and neuroblastoma.To explore the possible role of it in neuroblastoma in children,and to provide evidence for the diagnosis,treatment and prognosis of neuroblastoma.Methods:A total of 42 cases of paraffin embedded tissue sections of children with neuroblastoma were treated in the Department of Hematology of the Affiliated Hospital of Qiingdao University from January 2013 to December 2017.All specimens were obtained by surgical removal and biopsy.PV-6000 immunohistochemical method was used to stain the paraffin section specimens of 42 children with anti Nestin(10C2)antibody,and the expression of nestin in the above pathological specimens was detected.The children were grouped according to the general clinical situation of the children,the primary site of the tumor,the amplification of N-myc gene,the pathological type and the clinical stage.The children were divided into groups according to the level of peripheral blood neuron specific enolase(NSE)and serum ferritin(SF)at the initial diagnosis.Chi square test was used to analyze whether there was any difference in the expression of nestin between the above groups.Speraman rank correlation test was used to analyze the correlation between nestin expression and the above clinical characteristics.The survival time and survival rate of children with different nestin expression were analyzed by single factor survival curve(Kaplan-Meier),and the difference of survival time and survival rate between groups was analyzed by Log-Rank test.Cox proportional hazards regression model was used to carry out multivariate survival analysis.Results:(1)Of the 42 cases of newly diagnosed children,24 cases of nestin positive expression were positive,the positive expression rate was 57.14%,of which 14 cases were positive and 10 cases were positive,and the positive rate of high expression was33.33%.(2)The expression of Nestin in different clinical groups:?The primary site of the tumor was 32 cases of the primary peritoneum and 10cases of non primary tumor in the peritoneum,the positive expression rate of nestin was71.87%(23/32)and 10.00%(1/10)respectively.The difference was statistically significant(?~2=11.91,P<0.05),and the high expression rate of nestin positive was40.62%(13/32)and 10.00%(1/10),respectively.There was no statistical significance.(?~2=3.21,P>0.05).?Bone marrow infiltration:the positive expression rates of nestin were 92.85%(13/14)and 39.28%(11/28)in the bone marrow infiltration group and no bone marrow infiltration group.The difference was statistically significant(?~2=10.93,P<0.05),and the high expression rate of nestin positive was 64.28%(9/14)and 17.85%(5/28)respectively,and the difference was statistically significant(?~2=9.05,P<0.05).The high expression of nestin was positively correlated with bone marrow infiltration(r=0.71,P<0.05).?N-myc gene amplification:N-myc gene amplification group and N-myc gene non amplification group,the positive expression rate of nestin was 86.67%(13/15)and40.74%(11/27),the difference was statistically significant(?~2=8.30,P<0.05),and the high expression rate of nestin positive was 73.33%(11/15)and 11.11%(3/27),and the difference was statistically significant(?~2=14.11,P<0.05).The high expression of nestin was positively correlated with N-myc gene amplification(r=0.83,P<0.05).?Pathological type:the positive expression rate of nestin was 81.48%(22/27)and13.33%(2/15)in poor prognosis group and good prognosis group.The difference was statistically significant(?~2=18.28,P<0.05),and the high expression rate of nestin positive was 51.85%(14/27)and 0(0/15)respectively.The difference was statistically significant(?~2=9.45,P<0.05)..The high expression of nestin was positively correlated with the poor prognosis and pathological type(r=0.75,P<0.05).?Clinical stages:the positive expression rates of nestin in III-IV and I-II children were 70.58%(24/34)and 0(0/8)respectively,and the difference was statistically significant(?~2=13.17,P<0.05),and the high expression rate of nestin positive was41.17%(14/34)and 0(0/8),respectively(?~2=4.94,P<0.05),respectively.The high expression of in was positively correlated with clinical stage(r=0.74,P<0.05).?Risk degree:the positive rate of nestin was 80.00%(20/25)and 23.52%(4/17)in high risk group and low medium risk group.The difference was statistically significant(?~2=13.17,P<0.05),and the high expression rate of nestin positive was 56.00%(14/25)and 0(0/17)respectively,and the difference was statistically significant(?~2=11.87,P<0.05).High expression of nestin was positively correlated with risk(r=0.72,P<0.05).?Neuron specific enolase(NSE):NSE>100ng/ml group and NSE<100ng/ml group,the positive expression rate of nestin was 69.23%(18/26)and 37.50%(6/16)respectively.The difference was statistically significant(?~2=4.07,P<0.05),nestin positive expression rate was 50.00%(13/26)and 6.25%,respectively,and the difference was statistically significant Significance(?~2=6.67,P<0.05);high expression of nestin was positively correlated with NSE level(r=0.79,P<0.05).?Serum ferritin(SF):SF>150ng/L group and SF<150ng/L group,nestin positive expression rate was 75.00%(18/24)and 33.33%(6/18),the difference was statistically significant(?~2=4.28,P<0.05),nestin positive expression rate was 54.16%(13/24)and5.55%,respectively(?~2=8.85,P<0.05).High expression of nestin was positively correlated with SF level(r=0.44,P<0.05).(3)The relationship between the expression of single factor survival analysis nestin and the survival time,survival rate and prognosis of children:the median survival time of children with nestin positive expression was(26.86+3.79)months,the 1 year survival rate(OS)was(80.31+9.02)%,the 2 year survival rate(OS)was(51.92+12.94)%,and 3years survival rate(OS)was(22.22+11.27)%,and the median survival time of the nestin negative expression of the children was long(49.74+4.32)months,the 1 year survival rate(OS)was(91.03+6.12)%,the 2 year survival rate(OS)was(85.75+13.21)%,and the survival rate of 3 years(OS)was(68.66+18.67)%,and the difference in the group was statistically significant(?~2=8.75,P<0.05).(4)Multi factor survival analysis showed that nestin(HR=3.22,P<0.05),N-myc gene(HR=2.37,P<0.05)and NSE(HR=1.68,P<0.05)were independent prognostic factors for the specific survival of neuroblastoma,and the risk ratio of nestin was higher than that of N-myc gene and NSE.Conclusion:1.The positive expression rate of nestin in neuroblastoma was 57.14%,the positive rate was 33.33%,the positive low expression rate was 23.80%,and the high expression was obviously higher than the low expression,which could be used as an effective biological marker for the neuroblastoma.2.The positive expression of Nestin was positively related to the risk factors of N-myc gene amplification,and the survival time and survival rate of children with positive expression were significantly lower,suggesting that nestin can be used as one of the accurate criteria for judging the prognosis of children with neurocytoma.3.Nestin can be used as an independent risk factor affecting the prognosis of neuroblastoma,and lay the foundation for further study. |
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