Carcinoma-associated fibroblasts promote the stemness and chemoresistance of colorectal cancer by transferring exosomal lncRNA H19.
Carcinoma-associated fibroblasts promote the stemness and chemoresistance of colorectal cancer by transferring exosomal lncRNA H19.
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癌相关成纤维细胞通过转移外泌体lncRNA H19促进结直肠癌的干细胞性和化疗耐药性
作者：
Ren J;Ding L;Zhang D;Shi G;Xu Q;Shen S;Wang Y;Wang T;Hou Y
长链非编码rna （lncRNAs）参与包括结直肠癌（CRC）在内的多种肿瘤的病理过程。肿瘤微环境中癌相关成纤维细胞（CAFs）和癌细胞之间的串扰促进肿瘤的发展并赋予化疗耐药。在这项研究中，我们进一步研究了CAFs和参与这些过程的分子介质的潜在促肿瘤作用。方法：建立AOM/ dss诱导的结肠炎相关癌（CAC）小鼠模型，进行RNA测序。小干扰RNA （siRNA）序列被用来敲除H19。流式细胞术检测细胞凋亡。用稳定敲除H19的SW480细胞建立异种移植物模型。免疫组化法检测异种移植瘤组织中指示蛋白水平，TUNEL细胞凋亡法检测细胞凋亡。采用RNA-FISH和免疫荧光法检测H19在肿瘤间质和癌巢中的表达。AldeRed ALDH检测法检测细胞内醛脱氢酶（ALDH）活性。分离的外泌体通过透射电镜、纳米颗粒跟踪和Western blotting进行鉴定。结果：H19在CAC小鼠肿瘤组织中的表达高于正常结肠组织。在不同肿瘤淋巴结转移（TNM）阶段的CRC患者样本中也证实了H19的上调。此外，H19与结直肠癌标本中结直肠癌干细胞（CSCs）的干性相关。H19促进了CSCs的干性，增加了肿瘤启动细胞的频率。RNA-FISH显示H19在肿瘤基质中的表达高于癌巢。值得注意的是，H19在caf衍生的条件培养基和外泌体中富集，这反过来促进了CSCs的干性和CRC细胞的体外和体内化疗耐药。此外，H19作为CRC中miR-141的竞争内源性RNA海绵，激活了β-catenin通路，而miR-141显著抑制CRC细胞的干性。结论：CAFs通过转移外泌体H19促进结直肠癌的发生和耐药。H19通过作为miR-141的竞争内源性RNA海绵激活β-catenin通路，而miR-141抑制CRC细胞的干性。我们的研究结果表明，H19在结直肠肿瘤基质的cas中表达有助于肿瘤的发展和化疗耐药。
Long non-coding RNAs (lncRNAs) are involved in the pathology of various tumors, including colorectal cancer (CRC). The crosstalk between carcinoma- associated fibroblasts (CAFs) and cancer cells in the tumor microenvironment promotes tumor development and confers chemoresistance. In this study, we further investigated the underlying tumor-promoting roles of CAFs and the molecular mediators involved in these processes. Methods: The AOM/DSS-induced colitis-associated cancer (CAC) mouse model was established, and RNA sequencing was performed. Small interfering RNA (siRNA) sequences were used to knock down H19. Cell apoptosis was measured by flow cytometry. SW480 cells with H19 stably knocked down were used to establish a xenograft model. The indicated protein levels in xenograft tumor tissues were confirmed by immunohistochemistry assay, and cell apoptosis was analyzed by TUNEL apoptosis assay. RNA-FISH and immunofluorescence assays were performed to assess the expression of H19 in tumor stroma and cancer nests. The AldeRed ALDH detection assay was performed to detect intracellular aldehyde dehydrogenase (ALDH) enzyme activity. Isolated exosomes were identified by transmission electron microscopy, nanoparticle tracking and Western blotting. Results: H19 was highly expressed in the tumor tissues of CAC mice compared with the expression in normal colon tissues. The up-regulation of H19 was also confirmed in CRC patient samples at different tumor node metastasis (TNM) stages. Moreover, H19 was associated with the stemness of colorectal cancer stem cells (CSCs) in CRC specimens. H19 promoted the stemness of CSCs and increased the frequency of tumor-initiating cells. RNA-FISH showed higher expression of H19 in tumor stroma than in cancer nests. Of note, H19 was enriched in CAF-derived conditioned medium and exosomes, which in turn promoted the stemness of CSCs and the chemoresistance of CRC cells in vitro and in vivo. Furthermore, H19 activated the β-catenin pathway via acting as a competing endogenous RNA sponge for miR-141 in CRC, while miR-141 significantly inhibited the stemness of CRC cells. Conclusion: CAFs promote the stemness and chemoresistance of CRC by transferring exosomal H19. H19 activated the β-catenin pathway via acting as a competing endogenous RNA sponge for miR-141, while miR-141 inhibited the stemness of CRC cells. Our findings indicate that H19 expressed by CAFs of the colorectal tumor stroma contributes to tumor development and chemoresistance.
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影响因子：
8
作者：
通讯作者：
--
影响因子：
64.5
作者：
Boelens MC;Wu TJ;Nabet BY;Xu B;Qiu Y;Yoon T;Azzam DJ;Twyman-Saint Victor C;Wiemann BZ;Ishwaran H;Ter Brugge PJ;Jonkers J;Slingerland J;Minn AJ
通讯作者：
Minn AJ
影响因子：
64.5
作者：
GRODEN, J;THLIVERIS, A;WHITE, R
通讯作者：
WHITE, R
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.8
作者：
Gauck D;Keil S;Niggemann B;Zänker KS;Dittmar T
通讯作者：
Dittmar T