基于GPSM2的PDTX模型在胰腺癌个体化诊治中的应用

Application of PDTX Model Based on GPSM2 in Individualized Diagnosis and Treatment of Pancreatic Cancer

目的建立胰腺癌的人源肿瘤异种移植(patient derived tumor xenograft,PDTX)模型来验证G蛋白信号调控因子2(G-protein signaling modulator 2,GPSM2)作为治疗靶点的可行性。方法收集胰腺癌患者手术标本以建立PDTX模型,进行动物药敏实验,对使用治疗药物有效性做出评估。将实验小鼠分为对照组、sh-GPSM2组和5-氟尿嘧啶(fluorouracil,5-FU)化疗组。实验小鼠荷瘤取出、处理后,对不同化疗天数及相同天数不同化疗组的小鼠进行瘤体体积及重量统计分析。实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)检测sh-GPSM2组与对照组中GPSM2的表达水平。Western blot法检测3组小鼠肿瘤组织中ki-67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达水平。流式细胞术检测3组小鼠肿瘤细胞的凋亡和周期情况。结果成功建立PDTX模型,其中5-FU化疗组、sh-GPSM2组小鼠肿瘤瘤体相对于对照组有明显缩小趋势,PDTX模型肿瘤的生长被抑制(P<0.001)。RT-qPCR检测结果显示,sh-GPSM2组较对照组小鼠的GPSM2表达水平明显下调(P<0.001)。Western blot法检测结果显示,5-FU化疗组、sh-GPSM2组小鼠肿瘤组织中ki-67、PCNA表达水平相较于对照组明显降低(P<0.01)。流式细胞术分析结果显示,与对照组比较,5-FU化疗组和sh-GPSM2组小鼠的细胞在早期凋亡率升高,5-FU化疗组和sh-GPSM2组小鼠的细胞在细胞的S期百分比升高(P<0.05)。结论建立胰腺癌的PDTX模型应用于胰腺癌个体化诊疗,验证抑制GPSM2表达能够抑制PDTX模型胰腺癌的生长,sh-GPSM2制剂可能开发为一种新型的靶向药物。

Objective To establish the patient derived tumor xenograft(PDTX)model of pancreatic cancer and verify the feasibility of G-protein signaling modulator 2(GPSM2)as a therapeutic target.Methods Surgical specimens from pancreatic cancer patients were collected to establish PDTX model,and animal drug susceptibility experiments were conducted to evaluate the effectiveness of therapeutic drugs.The experimental mice were divided into three groups:control group,sh-GPSM2group,and 5-FU chemotherapy group.After the tumor-bearing mice were taken out and treated,the tumor volume and weight for mice with different chemotherapy days and different chemotherapy groups in the same days were statistically analyzed.The expression levels of GPSM2 in the sh-GPSM2group and the control group were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The expression levels of ki-67 and proliferating cell nuclear antigen(PCNA)in tumor tissues of the three groups were detected by Western blot.The apoptosis and cycle of tumor cells in the three groups were detected by flow cytometry.Results The PDTX model was successfully established.Compared with the control group,the tumor body of the sh-GPSM2group in the 5-FU chemotherapy group was significantly reduced,and the growth of the PDTX model tumor was inhibited(P<0.001).Compared with the control group,the results of RT-qPCR showed that the expression of GPSM2 in the sh-GPSM2group was significantly down-regulated(P<0.001).The results of Western blot test showed that the expression levels of ki-67 and PCNA in tumor tissues of 5-FU chemotherapy group and sh-GPSM2group were significantly lower than those of the control group(P<0.01).Flow cytometry analysis showed that compared with the control group,the apoptosis rate of cells in the 5-FU chemotherapy group and sh-GPSM2group increased in the early stage,and the percentage of cells in the S phase of the 5-FU chemotherapy group and sh-GPSM2group increased(P<0.001).Conclusion The established PDTX model of pancreatic cancer is applied to individualized diagnosis and treatment of pancreatic cancer,and it is verified that inhibiting the expression of GPSM2 can inhibit the growth of PDTX model pancreatic cancer.The sh-GPSM2 preparation may be developed as a new type of targeted drug.