Side population cells of pancreatic cancer show characteristics of cancer stem cells responsible for resistance and metastasis
Side population cells of pancreatic cancer show characteristics of cancer stem cells responsible for resistance and metastasis
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DOI：
10.1007/s11523-014-0323-z
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发表时间：
2015-06-01
影响因子：
5.4
通讯作者：
Bruns, Christiane J.
中科院分区：
文献类型：
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作者：
Niess, Hanno;Camaj, Peter;Bruns, Christiane J.
肿瘤干细胞（Cancer stem cells, CSCs）已被认为是实体肿瘤中肿瘤生长的起始和维持以及化疗耐药发展的基础。这些重要细胞在胰腺癌中的鉴定和作用仍然存在争议。在这里，我们从高度侵袭性和转移性的人胰腺癌细胞系L3.6pl中分离出侧群（SP）细胞，并评估它们作为CSCs模型的潜在作用。L3.6pl细胞采用Hoechst 33342染色分离SP细胞。将SP细胞、非SP细胞和未分选的L3.6pl细胞原位移植到裸鼠胰腺中，观察肿瘤生长情况。采用全基因组阵列分析RNA，绘制通路图。开发了L3.6pl的耐药变体，检测了SP比例，并评估了已知CSC标记的表面表达。从L3.6pl（0.9% +/- 0.22）中分离出具有自我更新和向非SP细胞分化能力的独特SP细胞。SP细胞在原位注射后表现出高度的致瘤性和转移性。转录组学分析发现，相对于非SP细胞，SP细胞中与肿瘤发生、分化和转移相关的基因网络调节。与肿瘤干细胞相关的Wnt、NOTCH和EGFR信号通路在SP细胞中发生改变。随着吉西他滨浓度的增加，SP细胞、ABCG2（+）和CD24（+）细胞的比例显著增加，而5-氟尿嘧啶（5-FU）处理降低了SP细胞的比例。SP细胞不同于先前假定的胰腺CSC标记物阳性细胞。L3.6pl细胞中hoechst诱导的侧群包括一个肿瘤细胞亚群，表现出侵袭性生长和转移，吉西他滨-增加，但不耐5-FU。这些细胞可以作为CSC生物学的部分模型。
Cancer stem cells (CSCs) have been proposed to underlie the initiation and maintenance of tumor growth and the development of chemoresistance in solid tumors. The identification and role of these important cells in pancreatic cancer remains controversial. Here, we isolate side population (SP) cells from the highly aggressive and metastatic human pancreatic cancer cell line L3.6pl and evaluate their potential role as models for CSCs. SP cells were isolated following Hoechst 33342 staining of L3.6pl cells. SP, non-SP, and unsorted L3.6pl cells were orthotopically xenografted into the pancreas of nude mice and tumor growth observed. RNA was analyzed by whole genome array and pathway mapping was performed. Drug resistant variants of L3.6pl were developed and examined for SP proportions and evaluated for surface expression of known CSC markers. A distinct SP with the ability to self-renew and differentiate into non-SP cells was isolated from L3.6pl (0.9 % +/- 0.22). SP cells showed highly tumorigenic and metastatic characteristics after orthotopic injection. Transcriptomic analysis identified modulation of gene networks linked to tumorigenesis, differentiation, and metastasization in SP cells relative to non-SP cells. Wnt, NOTCH, and EGFR signaling pathways associated with tumor stem cells were altered in SP cells. When cultured with increasing concentrations of gemcitabine, the proportion of SP cells, ABCG2(+), and CD24(+) cells were significantly enriched, whereas 5-fluorouracil (5-FU) treatment lowered the percentage of SP cells. SP cells were distinct from cells positive for previously postulated pancreatic CSC markers. The Hoechst-induced side population in L3.6pl cells comprises a subset of tumor cells displaying aggressive growth and metastasization, increased gemcitabine-, but not 5-FU resistance. The cells may act as a partial model for CSC biology.