Isolation and identification of chemotherapy-enriched sphere-forming cells from a patient with gastric cancer
Isolation and identification of chemotherapy-enriched sphere-forming cells from a patient with gastric cancer
复制标题
DOI：
10.1002/jcp.26627
复制
发表时间：
2018-10-01
影响因子：
5.6
通讯作者：
Abbaszadegan, Mohammad R.
中科院分区：
文献类型：
--
作者：
Bagheri, Vahid;Memar, Bahram;Abbaszadegan, Mohammad R.
胃癌(GC)分别是全球男性和女性癌症相关死亡的第三和第五位原因。尽管手术和化疗用于GC治疗，但到目前为止还没有有效的治疗方案。肿瘤干细胞(CSCs)在肿瘤的发生、生长、进展、侵袭、远处转移、复发和抗癌药物耐药等方面发挥着重要作用，是肿瘤治疗的重要靶点。在这里，我们从一名接受化疗的患者身上分离和鉴定了干细胞。组织消化后的小部分分离细胞在无血清条件下形成球状集落。这些球状集落在含血清的培养液中分化为上皮样细胞。少数球状细胞携带CD44和CD54标记，过表达与肿瘤生长和血管生成有关的DLL4。皮下注射不同传代的球状细胞可增强裸鼠的致瘤性。在体外和体内，球状细胞上调CD44基因CD44v3、-v6和-v8-10，干性因子OCT4、SOX2、SALL4和Cripto-1，自我更新分子IHH、Wnt、-catenin和BMI1，以及上皮间质转化(EMT)标记Twist1和Snail1。此外，这些细胞类似于从没有化疗的患者中分离出来的球状细胞，表达Oct-4和-catenin蛋白。然而，Twist1蛋白只在接受化疗的患者的球状细胞中表达。因此，这些细胞具有静止和迁移CSCs的所有特征，包括成瘤性、自我更新、多能性、侵袭和转移。综上所述，在GC患者中观察到的靶向化疗丰富的CSCs作为化疗耐药细胞可以提供比未经治疗的患者更有效的治疗策略。
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, -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 -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.