Sorting and identification of side population cells in the human cervical cancer cell line HeLa.
Sorting and identification of side population cells in the human cervical cancer cell line HeLa.
复制标题
人宫颈癌细胞HeLa侧群细胞的分选与鉴定
DOI：
10.1186/1475-2867-14-3
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发表时间：
2014-01-13
影响因子：
5.8
通讯作者：
Wei L
中科院分区：
文献类型：
--
作者：
Qi W;Zhao C;Zhao L;Liu N;Li X;Yu W;Wei L
多项报告显示，癌症干细胞（CSCs）存在于多种实体肿瘤中。一些研究表明，从不同癌细胞系中分离出的侧群（SP）细胞具有癌症干细胞样特性，但关于人类宫颈癌中SP细胞特性的研究报告却很少。本研究的目的是：1）找出一种检测宫颈癌中肿瘤干细胞样细胞的可行方法；2）分析分选得到的SP细胞的特性。
采用Hoechst 33342染色法和流式细胞术分析从人宫颈癌细胞系Hela中分离出SP细胞和非SP细胞。观察SP细胞和非SP细胞的形态。通过对SP细胞和非SP细胞进行免疫细胞化学研究，检测各种可能与癌症干细胞相关的生物标志物的表达。我们还对分选后的细胞进行了细胞周期和细胞凋亡分析。通过在非肥胖型糖尿病/重症联合免疫缺陷（NOD/SCID）小鼠体内形成肿瘤来分析SP细胞和非SP细胞的致瘤性。采用MTS法评估SP细胞、非SP细胞和Hela细胞之间的耐药性和抗辐射指数。
Hela细胞中SP细胞的比例约为1.07±0.32%。SP细胞比非SP细胞更小且更圆，大多呈集落样生长。免疫细胞化学显示，干细胞标志物（Oct3/4、CD133、BCRP）在SP细胞中高表达。此外，非SP细胞中的凋亡细胞数量（17.6±3.7%）明显高于SP细胞中的凋亡细胞数量（4.4±1.2%）。SP细胞在体内生长的肿瘤的HE染色显示分化更差，尽管在NOD/SCID小鼠的致瘤性方面SP细胞和非SP细胞之间没有显著差异。此外，与非SP细胞和Hela细胞相比，SP细胞对曲古抑菌素A（TSA）表现出更高的耐药性。还发现SP细胞对放射治疗更具抗性。
SP细胞具有一些癌症干细胞的特性，即高增殖能力、化学抗性和放射抗性，这可能有助于阐明未来宫颈癌有效临床治疗的新靶点。
Several reports have revealed that cancer stem cells (CSCs) exist in many types of solid tumors. Some studies have demonstrated that side population (SP) cells isolated from diverse cancer lines harbor cancer stem-like properties, but there are few reports examining the characteristic of SP cells in human cervical cancer. The aim of this study is 1) to find out a feasible way to detect the tumor stem-like cells in cervical cancer, and 2) to analyze the properties of the SP cells being sorted. Isolated SP and non-SP cells from human cervical cancer cell line Hela by Hoechst 33342 dying method and flow cytometry analysis. Observing morphology of SP and non-SP cells. The expression of various biomarkers putatively related to cancer stem cells were investigated by immucytochemistry of SP and non-SP cells. We also analyzed cell cycle and cell apoptosis for sorted cells. The oncogenicity of the SP and non-SP cells were analyzed by tumor formation in nonobesediabeti- c/severe combined immune- deficient (NOD/SCID) mice. The drug-resistant and radiation-resistant index between SP, non-SP and Hela cells was estimated by MTS assay. The fraction of SP cells in Hela was approximately 1.07 ± 0.32%. SP cells were smaller and rounder in shape than non-SP cells, and mostly showed colony-like growth. Immunocytochemistry showed that stem cell makers (Oct3/4, CD133, BCRP) were highly expressed in SP cells. Moreover, the number of apoptotic cells among non-SP cells (17.6 ± 3.7%) was significantly higher compared with that among SP cells (4.4 ± 1.2%). The HE staining of in vivo grown tumors result from SP cells showed more poor differentiation, though no significant differences were shown between SP and non-SP cells in NOD/SCID mice tumorigenicity. Furthermore, SP cells demonstrated a higher degree of drug resistance against trichostatin A (TSA) compared with that of non-SP and Hela cells. SP cells were also found to be more resistant against radiotherapy. SP cells possess some characteristics of CSCs, namely high proliferation ability, chemoresistance and radioresistance, which may be helpful to elucidate novel targets for effective clinical treatments of cervical cancer in the future.
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影响因子：
11.2
作者：
Kim, H;Lapointe, J;Pollack, JR
通讯作者：
Pollack, JR
影响因子：
5.2
作者：
Challen, GA;Little, MH
通讯作者：
Little, MH
影响因子：
4.7
作者：
Hu, Chen;Li, Hong;Gu, Jianren
通讯作者：
Gu, Jianren
影响因子：
3.5
作者：
Platet, Nadine;Mayol, Jean-Francois;Wion, Didier
通讯作者：
Wion, Didier
影响因子：
11.2
作者：
Li, Chenwei;Heidt, David G.;Simeone, Diane M.
通讯作者：
Simeone, Diane M.