TUG1和LCAL6长链非编码RNA的siRNA沉默对非小细胞肺癌患者来源异种移植瘤的影响

Effects of siRNA Silencing of TUG1 and LCAL6 Long Non-coding RNAs on Patient-derived Xenograft of Non-small Cell Lung Cancer.

作者信息

Fang Tian, Huang Hairong, Li Xiaoyou, Liao Jing, Yang Zhijian, Hoffman Robert M, Cheng X I, Liang Lei, Hu Wenjuan, Yun Shifeng

机构信息

Department of Comparative Medicine, Jinling Hospital, Clinical School of Medical College of Nanjing University, Nanjing, P.R. China.

Department of Cardiothoracic Surgery, Jinling Hospital, Clinical School of Medical College of Nanjing University, Nanjing, P.R. China.

出版信息

Anticancer Res. 2018 Jan;38(1):179-186. doi: 10.21873/anticanres.12206.

DOI:10.21873/anticanres.12206

PMID:29277771

Abstract

BACKGROUND/AIM: The aim of the present study was to establish a patient-derived xenograft (PDX) mouse model of non-small cell lung cancer (NSCLC) and investigate the anti-tumor efficacy of silencing of TUG1 and LCAL6 long non-coding RNA in the PDX model.

MATERIALS AND METHODS

PDXs were established by subcutaneously implanting NSCLC surgical tumor fragments into immunodeficient mice. PDX characterization was performed by histopathological, immunohistochemical and real-time polymerase chain reaction (RT-PCR) analyses for NSCLC subtype-specific markers and expression of LCAL6 and TUG1. Anti-tumor efficacy of siRNA silencing of TUG1 and LCAL6 was also investigated in the PDX model. The effect of TUG1 and LCAL6 silencing on protein expression of proliferation marker Ki67 and HOX-gene family HOXB7 in the tumors was assessed by immunohistochemical staining and Western blotting.

RESULTS

Establishment of NSCLC PDX models resulted in 9 of 26 cases (34.6%). Lung squamous cell carcinomas (SCC) had a higher engraftment rate (58.3%) than lung adenocarcinomas (ADC) (18.2%) (p<0.05). Comparative analysis indicated these established PDX models of NSCLC closely resembled the original tumors with regard to NSCLC subtype-specific markers TTF-1, napsin A, p63 and expression of LCAL6 and TUG1. The tumor volume and weight were significantly reduced in the TUG1-silenced group as compared to the control group (p<0.05). However, no significant tumor growth inhibition was found in the LCAL6-silenced group (p>0.05). Expression of both TUG1and LCAL6 was reduced by siRNA treatment. Expression of Ki67 and HOXB7 was significantly suppressed in both the TUG1- and LCAL6-silenced groups compared to the control group (p<0.01). The TUG1-silenced group showed more reduced Ki67 expression than the LCAL6-silenced group (p<0.05).

CONCLUSION

PDX NSCLC models were established with a high degree of similarity with the original tumor with regard to histological, immunohistochemical features and RNA expression of TUG1 and LCAL6. Silencing of TUG1 inhibited both tumor growth and expression of the proliferation marker Ki67 and HOX-gene family HOXB7 in the PDX model of NSCLC.

摘要

背景/目的：本研究旨在建立非小细胞肺癌（NSCLC）患者来源的异种移植（PDX）小鼠模型，并研究在该PDX模型中沉默TUG1和LCAL6长链非编码RNA的抗肿瘤疗效。

材料与方法

通过将NSCLC手术肿瘤碎片皮下植入免疫缺陷小鼠来建立PDX模型。通过组织病理学、免疫组织化学和实时聚合酶链反应（RT-PCR）分析对NSCLC亚型特异性标志物以及LCAL6和TUG1的表达进行PDX特征鉴定。还在PDX模型中研究了TUG1和LCAL6的siRNA沉默的抗肿瘤疗效。通过免疫组织化学染色和蛋白质印迹法评估TUG1和LCAL6沉默对肿瘤中增殖标志物Ki67和HOX基因家族HOXB7蛋白表达的影响。

结果

26例中有9例（34.6%）成功建立NSCLC PDX模型。肺鳞状细胞癌（SCC）的移植率（58.3%）高于肺腺癌（ADC）（18.2%）（p<0.05）。比较分析表明，这些建立的NSCLC PDX模型在NSCLC亚型特异性标志物TTF-1、napsin A、p63以及LCAL6和TUG1的表达方面与原始肿瘤非常相似。与对照组相比，TUG1沉默组的肿瘤体积和重量显著减小（p<0.05）。然而，在LCAL6沉默组中未发现明显的肿瘤生长抑制（p>0.05）。siRNA处理降低了TUG1和LCAL6的表达。与对照组相比，TUG1和LCAL6沉默组中Ki67和HOXB7的表达均显著受到抑制（p<0.01）。TUG1沉默组的Ki67表达比LCAL6沉默组降低得更多（p<0.05）。