Patel Shanaya, Shah Kavan, Mirza Sheefa, Shah Kanisha, Rawal Rakesh
Division of Medicinal Chemistry & Pharmacogenomics, Department of Cancer Biology, The Gujarat Cancer & Research Institute, Gujarat, India.
Department of Life Sciences, Gujarat University, Gujarat, India.
Oral Oncol. 2016 Nov;62:139-146. doi: 10.1016/j.oraloncology.2016.10.019. Epub 2016 Oct 27.
Circulating tumor cells (CTCs) are increasingly gaining importance due to their immense potential in enhancing diagnosis, prognosis and response to therapy in solid malignancies. Therefore, we aimed to comprehend the molecular diversity and critical role of this disseminated tumor population in OSCC.
CD44+ subpopulation was isolated using immuno-magnetic cell separation and their purity was validated using flow cytometry. Characterisation of self renewal potential and resistance to chemotherapy was assessed using tumor sphere forming and cytotoxicity assay. Gene expression profile of pertinent CSC (CD44s, CD44v3, CD44v6) and stemness markers (Bmi1 and Nanog) was carried out in CD44+ cells using Real Time PCR. Predominantly expressed markers and their association with clinico-pathological conditions were substantiated in 30 OSCC patients.
Flow cytometry analysis depicted a predominant population of CD44+CD24-CD45- cells suggesting that circulating tumor cells had a subpopulation of CSC like cells in the circulation. These cells demonstrated increased sphere forming capability and intrinsic chemo-resistance compared to non-CSC, thus indicating the CSC features of self-renewal and chemo-resistance. Additionally, CD44+ cells showed significantly increased expression levels of CD44v6 and Nanog compared to CD44- cells. Clinically, expression pattern of CD44v6 and Nanog correlated with different anatomical subsites, loco-regional aggressiveness of the disease and recurrence, thus opening newer avenues that can be explored for better prognostic and therapeutic implications.
This study explored the inevitable role of CD44v6 and Nanog as circulating stem like cell markers in assessment of loco-regional aggressiveness, detection of relapse and therapeutic response and resistance.
循环肿瘤细胞（CTCs）因其在实体恶性肿瘤的诊断、预后及治疗反应方面的巨大潜力而日益受到重视。因此，我们旨在了解这一播散性肿瘤群体在口腔鳞状细胞癌（OSCC）中的分子多样性及关键作用。
采用免疫磁珠细胞分选法分离CD44 +亚群，并通过流式细胞术验证其纯度。利用肿瘤球形成实验和细胞毒性实验评估自我更新潜力和对化疗的抗性。使用实时聚合酶链反应（Real Time PCR）检测CD44 +细胞中相关癌症干细胞（CSC）（CD44s、CD44v3、CD44v6）和干性标志物（Bmi1和Nanog）的基因表达谱。在30例OSCC患者中证实了主要表达的标志物及其与临床病理状况的关联。
流式细胞术分析显示CD44 + CD24 - CD45 -细胞占主要群体，表明循环肿瘤细胞在循环中有类似CSC的细胞亚群。与非CSC相比，这些细胞表现出更强的球形成能力和内在化疗抗性，从而表明其具有自我更新和化疗抗性的CSC特征。此外，与CD44 -细胞相比，CD44 +细胞中CD44v6和Nanog的表达水平显著升高。临床上，CD44v6和Nanog的表达模式与不同的解剖亚部位、疾病的局部区域侵袭性和复发相关，从而为更好的预后和治疗意义探索开辟了新途径。
本研究探讨了CD44v6和Nanog作为循环干细胞样细胞标志物在评估局部区域侵袭性、检测复发以及治疗反应和抗性方面的重要作用。
