失巢凋亡作为一种特殊的程序化细胞死亡形式,在机体发育、组织自身平衡、疾病发生和肿瘤转移中起重要作用。利用贴壁增殖表皮细胞系海拉(Hela)细胞作为模型,通过对比贴壁细胞悬浮后悬浮时间对细胞活性的影响,深入探讨了细胞失巢凋亡现象。研究考察了悬浮5min、30min、1h,2h,4h和8h后的活细胞数量,以及重新贴壁培养后对细胞活性(即贴壁24h后活细胞增殖数量)的影响。结果表明:悬浮时间可导致海拉细胞数量减少及活性降低,在细胞实验过程中应当严格控制悬浮细胞接种时长。
Abstract
Anoikis effect is a special program death for attachment cells in the body, which plays an important role in body development, tissue homeostasis, disease occurrence, and tumor metastasis. Attachment epithelial cell line Hela was hired as the experiment model to investigate the anoikis effect in detail. Living Hela cells were counted after trypsinization and floating for 5min, 30min, 1h, 2h, 4h, and 8h respectively. The activity of these cells was detected after seeding back for 24h. The results showed that floating time would severely decrease cell counting and activity of HeLa, and the floating time of attachment cell needs to be controlled during cell experiments.
关键词
计量学;生命科学计量 /
细胞数量 /
细胞活性 /
细胞测量 /
失巢凋亡 /
Hela细胞 /
悬浮时间
Key words
metrology /
life science metrology /
cell number /
cell activity /
cell measurement /
anoikis effect /
Hela cell /
floating time
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参考文献
［1］刘佳敏, 贾晓青, 狄斌. 三维细胞模型在肿瘤研究中的应用 ［J］. 中国药科大学学报, 2020, 51(2): 152-160.
Liu J M, Jia X Q, Di B. Application of three-dimensional cell model in tumor research ［J］. J China Pharm Univ, 2020, 51(2): 152-160.
［2］Wong C H, Siah K W, Lo A W. Estimation of clinical trial success rates and related parameters ［J］. Biostatistics, 2019, 20(2): 273-286.
［3］Amelian A, Wasilewska K, Megias D, et al. Application of standard cell cultures and 3D in vitro tissue models as an effective tool in drug design and development ［J］. Pharmacological Reports, 2017, 69(5): 861-870.
［4］Faruqui N, Kummrow A, Fu B, et al. Cellular Metrology: Scoping for a Value Proposition in Extra-and Intracellular Measurements ［J］. Front Bioeng Biotechnol, 2020, 7: 1-8.
［5］金力. 计量测试赋能生命科学高质量发展［J］. 质量与标准化, 2021(6): 1-4.
Jin L. Quantitative testing enables high-quality development of life science ［J］. Quality and Standardization, 2021(6): 1-4.
［6］李晶晶, 罗明志, 杨莹,等. 三种细胞计数法的对比试验研究 ［J］. 江西科技师范大学学报, 2016(6):79-81.
Li J J, Luo M Z, Yang Y, et al. Comparison of Three Cell Count Methods ［J］. Journal of Jiangxi Science & Technology Normal University, 2016(6): 79-81.
［7］Scherer W F, Syverton J T, Gey G O. Studies on the propagation in vitro of poliomyelitis viruses. IV. Viral multiplication in a stable strain of human malignant epithelial cells (strain HeLa) derived from an epidermoid carcinoma of the cervix ［J］. J Exp Med, 1953, 97(5): 695-710.
［8］Mirabelli P, Coppola L, Salvatore M. Cancer Cell Lines Are Useful Model Systems for Medical Research ［J］. Cancers (Basel), 2019 , 11(1098): 1-18.
［9］薛志超, 曾嘉明,李永淑, 等. 接种数量对细胞生长和药物作用的影响及细胞计数方法对比［J］. 计量学报, 2023, 44(3): 341-349.
Xue Z C, Zeng J M, Li Y S, et al. Cell Growth and Drug Effect Differences Induced by Inoculum Number Variations and Comparison of Cell Counting Methods［J］. Acta Metrologica Sinica, 2023, 44(3): 341-349.
［10］Paoli P, Giannoni E, Chiarugi P. Anoikis Molecular Pathways and Its Role in Cancer Progression ［J］. Biochim Biophys Acta, 2013, 1833(12): 3481-3498.
［11］Cai Q, Yan L, Xu Y. Anoikis Resistance Is a Critical Feature of Highly Aggressive Ovarian Cancer Cells ［J］. Oncogene, 2014, 34(25): 3315-3324.
［12］Peppicelli S, Ruzzolini J, Bianchini F, et al. Anoikis Resistance as a Further Trait of Acidic-adapted Melanoma Cells ［J］. Journal of Oncology, 2019(25): 1-13.
［13］Kayo H, Hayate Y, Kouhei K, et al. Cyclin A is a reliable proliferation marker in endometrial cancer cell lines ［J］. 2019, 17(5): 4455-4462.
［14］Desdouets C, Sobczak-thépot J, Murphy M, et al. Cyclin A: function and expression during cell proliferation ［J］. Prog Cell Cycle Res, 1995(1): 115-123.
［15］Wu Y, Wu M, He G, et al. Glyceraldehyde-3-phosphate Dehydrogenase: a Universal Internal Control for Western Blots in Prokaryotic and Eukaryotic Cells［J］. Anal Biochem, 2012, 423(1): 15-22.
［16］Akekawatchai C, Roytrakul S, Phaonakrop N, et al. Proteomic Analysis of the Anoikis-resistant Human Breast Cancer Cell Lines ［J］. Methods Mol Biol, 2020, 2138: 185-193
［17］Chang J C. Cancer Stem Cells: Role in Tumor Growth, Recurrence, Metastasis, and Treatment Resistance ［J］. Medicine (Baltimore), 2016, 95(Suppl 1): 20-25.
［18］Huyen N T B, Prachayasittikul V, Chan-on W. Anoikis-resistant Cholangiocarcinoma Cells Display Aggressive Characteristics and Increase Stat3 Activation. ［J］. J Hepatobiliary Pancreat Sci, 2016, 23(7): 397-405.
基金
中国计量科学研究院院基本科研业务费(AKYZZ2224)
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