Isolation and identification of chemotherapy‐enriched sphere‐forming cells from a patient with gastric cancer

Abstract

Gastric cancer (GC) is the third and fifth cause of cancer-associated mortality for men and women throughout the world, respectively. Despite the use of surgery and chemotherapy for GC therapy, there are no efficient therapeutic protocols for it to date. Cancer stem cells (CSCs) due to their pivotal role in tumor initiation, growth, progression, invasion, distant metastasis, recurrence and resistance to anticancer drugs are very appealing targets for cancer therapies. Here, we isolated and identified CSCs from a chemotherapy-treated patient. Small subpopulation of dissociated cells after tissue digestion formed spheroid colonies in serum-free media under the non-adherent condition. These spheroid colonies differentiated into epithelial like cells in serum-containing medium. Few sphere-forming cells carried CD44 and CD54 markers overexpressed DLL4 that is responsible for tumor growth and angiogenesis. Subcutaneous injections of sphere-forming cells in different passages conferred tumorigenicity in nude mice. Sphere-forming cells upregulated CD44 polymorphisms CD44v3, -v6, and -v8 -10, stemness factors OCT4, SOX2, SALL4 and Cripto-1, self-renewal molecules IHh, Wnt, β-catenin and BMI1, and epithelial mesenchymal transition (EMT) markers Twist1 and Snail1 in vitro and in vivo. Moreover, these cells similar to sphere-forming cells isolated from a chemotherapy-free patient expressed Oct-4 and β-catenin proteins. However, the Twist1 protein was only expressed by sphere-forming cells derived from the chemotherapy-treated patient. Thus, these cells have all the characteristics of stationary and migratory CSCs, including tumorigenicity, self-renewal, pluripotency, invasion and metastasis. Taken together, targeting chemotherapy-enriched CSCs as chemo-resistance cells observed in GC patients can provide more effective therapeutic strategies compared to untreated patients.

Synergistic antitumor activity of a DLL4/VEGF bispecific therapeutic antibody in combination with irinotecan in gastric cancer

“…However, both anti-hDLL4 and ABL001 had a significant effect on AGS and MKN-28 cells in 3D-culture reducing sphere formation (indicative of stem-like cells). CSCs are responsible for tumor relapse, chemoresistance, and cancer metastasis ( 8 , 27 ). Conventional chemotherapy offers limited benefits in advanced-stage GC patients and prognosis for such patients remains poor.…”

“…In mammals, there are four Notch homologous receptors (Notch 1-4), which can bind various ligands–Delta-like ligands (DLL1, 3, and 4), and Jagged 1 and 2 ( 7 ). DLL4, one of the major factors involved in Notch signaling, was shown to be responsible for tumor formation, EMT, and self-renewal as CSCs ( 8 ). Targeting DLL4 expression may be a feasible approach to interfere with angiogenesis, tumorigenesis, and metastasis for anti-cancer effect ( 9 , 10 ).…”

Histone deacetylases inhibitor MS‐275 suppresses human esophageal squamous cell carcinoma cell growth and progression via the PI3K/Akt/mTOR pathway

“…Many studies indicate that tumors contain a small subset of cancer stem cells (CSCs). Sphere‐forming cells have CSCs properties, which are more resistant to chemotherapy and more tumorigenic compared with common tumor cells (Bagheri et al, ; Cao et al, ). We examined the ability of KYSE‐70 and EC9706 to form spheres in the presence of MS‐275.…”

“…Most importantly, HDAC1, HDAC2, HDAC3 (Ahrens et al, 2015;Koumangoye et al, 2015;Li et al, 2017;Zhong et al, 2018), and HDAC4 (Zeng et al, 2016) Many studies indicate that tumors contain a small subset of cancer stem cells (CSCs). Sphere-forming cells have CSCs properties, which are more resistant to chemotherapy and more tumorigenic compared with common tumor cells (Bagheri et al, 2018;Cao et al, 2011 EMT is a process that epithelial cancer cells transform to facilitate the loss of epithelial features and gain of mesenchymal phenotype (Xu et al, 2018). EMT promotes cancer cell progression and metastasis, and is involved in the regulation of CSCs (Hou et al, 2016;van Zijl et al, 2009;Xu et al, 2018).…”

https://researchopenworld.com/development-of-synthetic-cell-differentiation-agent-formulations-for-the-prevention-and-therapy-of-cancer-via-targeting-of-cancer-stem-cells/#

“…Aberrant DNA methylations happen more readily when DNA synthesis is slowing down by cytotoxic drugs or radiation [113]. Another benefit to implement destabilization of abnormal methylation enzymes is to avoid expansion of CSC population induced by the destruction of the tumor caused by cytotoxic chemotherapy [114], or due to tumor progression [103][104][105][106]. Reconstruction of the tumor is a major mission of CSCs.…”