TM4SF1 Promotes Gemcitabine Resistance of Pancreatic Cancer In Vitro and In Vivo.
TM4SF1 Promotes Gemcitabine Resistance of Pancreatic Cancer In Vitro and In Vivo.
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TM4SF1 促进胰腺癌体外和体内吉西他滨耐药
DOI：
10.1371/journal.pone.0144969
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Logsdon CD
中科院分区：
文献类型：
--
作者：
Cao J;Yang J;Ramachandran V;Arumugam T;Deng D;Li Z;Xu L;Logsdon CD
背景TM 4SF 1在胰腺导管腺癌（PDAC）中过表达，并影响这种癌症的发展。此外，多药耐药（MDR）通常与胰腺癌中的肿瘤化疗耐药性相关。然而，TM 4SF 1与MDR的相关性尚不清楚。本研究旨在探讨TM 4SF 1在PDAC吉西他滨耐药中的作用，并探讨其与MDR的可能分子机制。方法采用定量RT-PCR方法检测胰腺癌细胞系和人胰腺导管上皮细胞系中TM 4SF 1的表达。使用Hiperfect转染试剂进行TM 4SF 1 siRNA转染以敲低TM 4SF 1。采用RT-PCR、RT-PCR和Western blotting等方法对转录本进行分析。体外实验观察沉默TM 4SF 1后胰腺癌细胞对吉西他滨的敏感性。我们证明了TM 4SF 1的细胞信号介导的癌细胞的化疗耐药性，通过评估多药耐药（MDR）基因的表达，使用定量RT-PCR。在体内，我们使用原位胰腺肿瘤模型来研究在MIA PaCa-2细胞系中通过慢病毒介导的shRNA沉默TM 4SF 1后对增殖的影响。结果7种胰腺癌细胞系中TM 4SF 1 mRNA的表达均高于HPDE细胞系。TM 4SF 1在3个吉西他滨敏感细胞系（L3.6pl、BxPC-3、SU86.86）中的表达低于4个吉西他滨耐药细胞系（MIA PaCa-2、PANC-1、Hs 766 T、AsPC-1）。我们评估了TM 4SF 1是胰腺癌细胞吉西他滨耐药的假定靶点。使用AsPC-1、MIA PaCa-2和PANC-1，我们研究了TM 4SF 1沉默影响细胞增殖，并与阴性对照处理的细胞相比，增加吉西他滨处理介导的细胞凋亡百分比。这种耐药性与多药耐药基因ABCB 1和ABCC 1的表达有关。在体内，MIA PaCa-2细胞系中TM 4SF 1的沉默增加了使用非侵入性生物发光成像评估的原位胰腺肿瘤模型中基于吉西他滨的治疗的有效性。结论TM 4SF 1是胰腺癌细胞高表达的表面膜抗原，可增加胰腺癌细胞对吉西他滨的耐药性。因此，TM 4SF 1可能是克服胰腺癌化疗耐药性的一个有希望的靶点。
Background TM4SF1 is overexpressed in pancreatic ductal adenocarcinoma (PDAC) and affects the development of this cancer. Also, multidrug resistance (MDR) is generally associated with tumor chemoresistance in pancreatic cancer. However, the correlation between TM4SF1 and MDR remains unknown. This research aims to investigate the effect of TM4SF1 on gemcitabine resistance in PDAC and explore the possible molecular mechanism between TM4SF1 and MDR. Methods The expression of TM4SF1 was evaluated in pancreatic cancer cell lines and human pancreatic duct epithelial (HPDE) cell lines by quantitative RT-PCR. TM4SF1 siRNA transfection was carried out using Hiperfect transfection reagent to knock down TM4SF1. The transcripts were analyzed by quantitative RT-PCR, RT-PCR and western blotting for further study. The cell proliferation and apoptosis were obtained to investigate the sensitivity to gemcitabine of pancreatic cancer cells after silencing TM4SF1 in vitro. We demonstrated that cell signaling of TM4SF1 mediated chemoresistance in cancer cells by assessing the expression of multidrug resistance (MDR) genes using quantitative RT-PCR. In vivo, we used orthotopic pancreatic tumor models to investigate the effect of proliferation after silencing TM4SF1 by a lentivirus-mediated shRNA in MIA PaCa-2 cell lines. Results The mRNA expression of TM4SF1 was higher in seven pancreatic cancer cell lines than in HPDE cell lines. In three gemcitabine-sensitive cell lines (L3.6pl, BxPC-3, SU86.86), the expression of TM4SF1 was lower than that in four gemcitabine-resistant cell lines (MIA PaCa-2, PANC-1, Hs766T, AsPC-1). We evaluated that TM4SF1 was a putative target for gemcitabine resistance in pancreatic cancer cells. Using AsPC-1, MIA PaCa-2 and PANC-1, we investigated that TM4SF1 silencing affected cell proliferation and increased the percentages of cell apoptosis mediated by treatment with gemcitabine compared with cells which were treated with negative control. This resistance was associated with the expression of multidrug resistance genes including ABCB1 and ABCC1. In vivo, silencing of TM4SF1 in MIA PaCa-2 cell lines increased the effectiveness of gemcitabine-based treatment in orthotopic pancreatic tumor models evaluated using noninvasive bioluminescent imaging. Conclusion These findings suggest that TM4SF1 is a surface membrane antigen that is highly expressed in pancreatic cancer cells and increases the chemoresistance to gemcitabine. Thus, TM4SF1 may be a promising target to overcome the chemoresistance of pancreatic cancer.