Effect Of X-Ray Irradiation Dose On Proliferation And Apoptosis Of Keloid Fibroblasts And Analysis Of The Clinical Efficacy Of Postoperative Adjuvant Radiotherapy For Keloid

Objective:To explore the effects of X-ray irradiation dose on the proliferation activity,late apoptosis ratio,expression of apoptosis genes and related functional proteins of keloid fibroblasts,and to evaluate the clinical efficacy of iridium-192 radiotherapy after keloid surgery.Method:1.Keloid dermis fibroblasts were cultured,and the cells in logarithmic growth phase were irradiated with 9/12/15 Gy X-rays.For 5 consecutive days,OD values were detected by enzyme labeling to reflect the proliferation of cells calibrated by CCK-8 kit for 1 hour.Flow cytometry was used to detect apoptosis labeled by Annexin V-FITC/PI apoptosis kit.Western blotting(WB)was used to detect the expression of related proteins(Bax,Bcl-2,TGF-β,VEGFA,β-Catenin),and ImageLab software was used to quantify the protein expression.IBM SPSS Statistics 25 statistical analysis software was used,the value was expressed by mean ±standard deviation,and the analysis method was one way ANOVA.2.Collect the keloid cases of clinical postoperative adjuvant radiotherapy,establish the patient database,follow up the curative effect of the patients on time and collect pictures of the affected area,and evaluate the curative effect by scoring the color,vascular distribution,thickness and softness of the affected area according to the Vancouver scar scale(VSS:Vancouver Scar Scale).Result:1.The keloid fibroblasts treated with radiotherapy dose(9/12/15 Gy)showed that the higher the total radiation dose,the stronger the effect.The proliferation inhibition rates of 9Gy,12Gy and 15Gy groups were 26%,31.6%and 32.8%,respectively.The late apoptosis rates of 0 Gy,9 Gy,12Gy and 15 Gy groups were 3.65%,4.86%,7.18%and 8.51%,respectively.The expression of Bax protein in 0 Gy,9 Gy,12 Gy and 15 Gy groups were "1",0.93,1.41 and 1.70,respectively.The expression of Bcl-2 protein in 0 Gy,9 Gy,12 Gy and 15 Gy groups was "1",0.55,0.52,0.29,respectively.The expression of VEGFA(vascular endothelial growth factor A)protein in 0 Gy,9 Gy,12 Gy and 15Gy groups were "1",0.68,0.68,0.70,respectively.The expression of TGF-β(transforming growth factor β)protein in 0 Gy,9 Gy,12 Gy and 15 Gy groups were "1",0.78,0.72,0.72,respectively.The expression ofβ-catenin(β-catenin)protein in 0Gy,9 Gy,12 Gy and 15Gy groups were "1",1.03,1.01,0.96,respectively.2.The average VSS score of 9 Gy group,12 Gy group and 15Gy group were 4.2,3.5,2.7respectively during clinical follow-up.The cure rate of the 9Gy group was 71%,the effective rate was 87.1%;the cure rate of the 12Gy group was 78.3%,the effective rate was 87%;the cure rate of the 15Gy group was 88.9%,and the effective rate was 100%.Conclusion:1.X-ray irradiation dose(9/12/15Gy)can inhibit the proliferation of keloid fibroblasts,increase the late apoptosis rate,activate apoptosis gene Bax and inhibit the expression of anti-apoptosis gene Bcl-2.The higher the total dose,the stronger the effect.2.Postoperative radiotherapy for keloid,the higher the total dose,the lower the scar score and the less the number of recurrence.Therefore,postoperative adjuvant radiotherapy for keloid is effective in reducing recurrence,and properly increasing the clinical radiation dose can lead to a higher cure rate.