Dclk1, a tumor stem cell marker, regulates pro-survival signaling and self-renewal of intestinal tumor cells

被引：97

作者：

Chandrakesan, Parthasarathy ^{[1
,2
,3
]}

Yao, Jiannan ^{[1
]}

Qu, Dongfeng ^{[1
,2
,3
]}

May, Randal ^{[1
,3
]}

Weygant, Nathaniel ^{[1
]}

Ge, Yang ^{[1
,7
]}

Ali, Naushad ^{[1
,3
]}

Sureban, Sripathi M. ^{[1
,3
]}

Gude, Modhi ^{[1
,4
]}

Vega, Kenneth ^{[1
,5
]}

Bannerman-Menson, Eddie ^{[8
]}

Xia, Lijun ^{[6
]}

Bronze, Michael ^{[1
]}

An, Guangyu ^{[7
]}

Houchen, Courtney W. ^{[1
,2
,3
,8
]}

机构：

[1] Univ Oklahoma, Hlth Sci Ctr, Dept Med, Div Digest Dis & Nutr, Oklahoma City, OK 73104 USA

[2] Univ Oklahoma, Hlth Sci Ctr, OU Canc Inst, Oklahoma City, OK USA

[3] Dept Vet Affairs Med Ctr, Oklahoma City, OK USA

[4] Access Endocrine Diabet & Thyroid Ctr, Oklahoma City, OK USA

[5] Natl Jewish Hlth, Dept Med, Denver, CO USA

[6] Oklahoma Med Res Fdn, 825 NE 13th St, Oklahoma City, OK 73104 USA

[7] Capital Med Univ, Beijing Chao Yang Hosp, Dept Oncol, Beijing, Peoples R China

[8] COARE Biotechnol, Oklahoma City, OK USA

来源：

MOLECULAR CANCER | 2017年 / 16卷

关键词：

Dclk1; APC mutation; Cancer stem cells; Intestinal epithelial cells; Pro-survival signaling; Self-renewal; Nanoparticles; Colorectal cancer; EPITHELIAL-MESENCHYMAL TRANSITION; MICRORNA-DEPENDENT MECHANISM; COLORECTAL-CANCER; TUFT CELLS; TRANSDUCTION PATHWAYS; BACTERIAL-INFECTION; PANCREATIC-CANCER; CAM KINASE-LIKE-1; XENOGRAFT GROWTH; NEOPLASIA MICE;

D O I：

10.1186/s12943-017-0594-y

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: More than 80% of intestinal neoplasia is associated with the adenomatous polyposis coli (APC) mutation. Doublecortin-like kinase 1 (Dclk1), a kinase protein, is overexpressed in colorectal cancer and specifically marks tumor stem cells (TSCs) that self-renew and increased the tumor progeny in Apc(Min/+) mice. However, the role of Dclk1 expression and its contribution to regulating pro-survival signaling for tumor progression in Apc mutant cancer is poorly understood. Methods: We analyzed DCLK1 and pro-survival signaling gene expression datasets of 329 specimens from TCGA Colon Adenocarcinoma Cancer Data. The network of DCLK1 and pro-survival signaling was analyzed utilizing the GeneMANIA database. We examined the expression levels of Dclk1 and other stem cell-associated markers, pro-survival signaling pathways, cell self-renewal in the isolated intestinal epithelial cells of Apc(Min/+) mice with high-grade dysplasia and adenocarcinoma. To determine the functional role of Dclk1 for tumor progression, we knocked down Dclk1 and determined the pro-survival signaling pathways and stemness. We used siRNA technology to gene silence pro-survival signaling in colon cancer cells in vitro. We utilized FACS, IHC, western blot, RT-PCR, and clonogenic (self-renewal) assays. Results: We found a correlation between DCLK1 and pro-survival signaling expression. The expression of Dclk1 and stem cell-associated markers Lgr5, Bmi1, and Musashi1 were significantly higher in the intestinal epithelial cells of Apc(Min/+) mice than in wild-type controls. Intestinal epithelial cells of Apc(Min/+) mice showed increased expression of pro-survival signaling, pluripotency and self-renewal ability. Furthermore, the enteroids formed from the intestinal Dclk1(+) cells of Apc(Min/+) mice display higher pluripotency and pro-survival signaling. Dclk1 knockdown in Apc(Min/+) mice attenuates intestinal adenomas and adenocarcinoma, and decreases pro-survival signaling and self-renewal. Knocking down RELA and NOTCH1 pro-survival signaling and DCLK1 in HT29 and DLD1 colon cancer cells in vitro reduced the tumor cells' ability to self-renew and survive. Conclusion: Our results indicate that Dclk1 is essential in advancing intestinal tumorigenesis. Knocking down Dclk1 decreases tumor stemness and progression and is thus predicted to regulate pro-survival signaling and tumor cell pluripotency. This study provides a strong rationale to target Dclk1 as a treatment strategy for colorectal cancer.

引用

页数：14