| Objective: A new fluorescent targeted probe was made of Glypican-3(GPC3)antibody and IRDye800 cw fluorescent dye.The fluorescence imaging of human liver cancer tissue in vitro was carried out to investigate the detection efficiency of near-infrared fluorescence imaging technology on the expression of GPC3 in liver cancer and the relationship between fluorescence intensity and histological characteristics of hepatocellular carcinoma,surgical prognosis was also analysed in this study.Using near-infrared fluorescence imaging technology with near-infrared fluorescence GPC3-target probe is a novel method to detect the expression of GPC3,which provides a new reference for the surgical treatment and postoperative treatment of hepatocellular carcinoma.Methods: 117 patients with liver cancer who underwent hepatectomy in the Affiliated Hospital of Southwest Medical University were selected.The GPC3-IRDye800 targeted probe was used for fluorescence imaging of tumor samples in vitro.The immersion concentration was 10 μ g / ml,the immersion dose was 10 ml,and the immersion time was 5min.After immersion,the unbound probe was washed with 5-10 ml fetal bovine blood,and then washed with phosphate buffer solution.After cleaning,the near-infrared fluorescence images of different kinds of liver cancers were obtained by near-infrared fluorescence technology.The expression of GPC3 in liver cancer was detected by Envision immunohistochemistry approach.The relationship between the expression of GPC3 and the intensity of fluorescence imaging,and the relationship between the intensity of fluorescence and the differentiation of HCC were analyzed.According to the fluorescence intensity of GPC3 targeted probe,HCC postoperative patients were divided into high fluorescence intensity group and low fluorescence intensity group.The progression free survival rate and overall survival rate of the two groups were analyzed by Kaplan Meier method.Univariate and multivariate Cox proportional hazard regression models were used to analyze the predictive value of GPC3 probe fluorescence intensity for progression free survival(PFS)and overall survival in HCC patients.Meanwhile,HCC patients were divided into AFP positive group(> 10 ng / ml)and AFP negative group(≤ 10 ng / ml).Kaplan-Meier method was used to analyze the difference of progression free survival rate between the low fluorescence intensity group and high fluorescence intensity group in AFP positive group and negative group respectively.Results: Among 117 cases of liver cancer,87 cases were hepatocellular carcinoma,20 cases were cholangiocarcinoma and 10 cases were metastatic carcinoma.Immunohistochemical results showed that the expression rate of GPC3 in liver cancers was 59.83%(70 / 117).A total of 70 cases of HCC tissues expressed GPC3,the expression rate was 80.46%(70 / 87).According to the GPC3 staining score,the high expression rate of GPC3 in HCC tissues was 66.67%(58 / 87).The results of fluorescence imaging showed that among69 positive samples,68 samples were positive for GPC3 in immunohistochemistry and 1 sample was negative for GPC3;among 48 samples,46 samples were negative for GPC3 and 2 samples were positive for GPC3.The sensitivity and specificity of GPC3 fluorescent probe were 97.14%(68 / 70)and 97.87%(46 / 47)respectively.The results of GPC3 detected by fluorescence imaging and immunohistochemistry were basically consistent(kappa = 0.964,P < 0.01),and there was no significant difference between these two methods(P > 0.05).Quantitative analysis of fluorescence intensity showed that the expression of GPC3 was proportional to the fluorescence intensity(P < 0.05).The progression free survival of patients with high fluorescence intensity of GPC3 probe was inferior than that of patients with low fluorescence intensity(7 months vs 13 months,P < 0.05).The one-year survival rate of GPC3 probe high fluorescence intensity group and low fluorescence group was 34.8% and 63.4% respectively,and the difference was statistically significant(P < 0.05).Multivariate analysis confirmed that GPC3 probe fluorescence intensity,microvascular invasion and differentiation were independent risk factors for progression free survival.In HCC patients with AFP negative,the fluorescence intensity of GPC3 probe was inversely proportional to the progression free survival(P < 0.05).Conclusion:The fluorescence intensity of GPC3 probe can also be used as an independent prognosis predictive factor of HCC,and the fluorescence intensity of GPC3 probe is inversely proportional to the progression free survival rate and overall survival rate of patients with HCC;in HCC patients with negative serum AFP,the fluorescence intensity of GPC3 probe has a certain predictive potential for prognosis... |
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