BioVector® NCI-N417 人小细胞肺癌细胞株

BioVector® NCI-N417 Human Small Cell Lung Carcinoma Cell Line

第一部分 中文说明

一 产品基本信息与遗传学背景

• 细胞名称 BioVector® NCI-N417 人小细胞肺癌细胞

• 别名与同义词 NCI-N-417、NCI-H417、N417、H417、NCIN417

• 系统学 Accession Cellosaurus CVCL_1602

• 物种来源 人类 Homo sapiens

• 组织与疾病背景 该细胞株由美国国家癌症研究所（NCI）的 Gazdar 及其合作者建立，源自一名确诊为肺小细胞癌（SCLC）患者的病灶，是研究经典神经内分泌型肺癌的标志性体外模型之一。

• 突变特征与基因组状态

  • 带有明确的 TP53 基因纯合/半合子突变（p.Glu298Ter，导致蛋白翻译提前终止）。

  • 带有 PIK3CA 基因杂合突变（p.Gln546Lys）。

  • 染色体易位 包含 PVT1-CLVS1 基因融合。

  • 特殊特征 包含集成的异种小鼠白血病病毒相关病毒（XMRV）Bxv-1，提示其在早期建立过程中可能经过了裸鼠异种移植阶段。

• 生物安全级别 1级（或根据其携带的内源反转录病毒片段成分，遵循常规 2级生物安全防护规范操作更为稳妥）。

二 细胞形态学与培养环境

• 形态学特征 展现典型的小细胞肺癌变异亚型（Variant subclass）形态特征。细胞体积相对较小，核质比极高。它们在培养基中极少贴壁，主要以松散的悬浮细胞团块、密集聚集体形式在液体中漂浮生长。

• 生长模式 悬浮聚集生长。

• 倍增时间 增殖较快，倍增时间大约为 26小时。

• 标准完全培养基配方

  • 基础培养基 BioVector® RPMI-1640 培养基。

  • 维持添加 10% BioVector® 优质胎牛血清（对于状态不佳或新复苏的细胞，可选用 15% 胎牛血清以加速恢复）。

• 物理培养参数 37摄氏度恒温、5% 二氧化碳、空气饱和湿度。

三 细胞传代与复苏标准操作步骤

1. 常规传代操作

   • 当悬浮的细胞簇生长得过于庞大、内部开始出现发黑的多重细胞重叠坏死，或者培养基显著变黄时需要进行传代。

   • 将包含细胞团的悬液转移至离心管中，以 150 g 离心 5分钟，弃去旧培养基。

   • 用新鲜的 BioVector® 完全培养基重悬细胞。传代时必须使用移液枪非常轻柔地吹打几次，将过大的团块打散成大小适中的小细胞丛。切勿使用机械手段强行彻底打散成绝对的单细胞，否则会破坏细胞间的集聚信号导致大批细胞死亡。

   • 推荐接种密度 维持在每毫升 2.0乘以10的5次方 至 1.0乘以10的6次方 个活细胞。传代比例通常为 1比2 至 1比4。

2. 冻存细胞复苏

   • 从液氮中取出冷冻管，立即投入 37摄氏度 BioVector® 水浴锅中快速摇动使其融化，控制在 1到2分钟内。

   • 将解冻的细胞悬液移至含 5到8 mL 预热培养基的离心管中，以 150 g 离心 5分钟。

   • 弃去含有二甲基亚砜（DMSO）的旧上清，加入新鲜 BioVector® 完全培养基重悬，接种到培养瓶内，静置培养。

四 核心科研应用方向

1. 小细胞肺癌（SCLC）耐药株的创制与逆转研究：BioVector® NCI-N417 广泛用于构建抗顺铂（Cisplatin-resistant, 如 N417/CDDP 细胞）和抗紫杉醇（Paclitaxel）的耐药模型，用于筛选多药耐药相关蛋白（MRP）表达和拓扑异构酶突变引起的靶向脱靶现象。

2. 肿瘤干细胞与缺氧微环境研究：该细胞在缺氧（Hypoxia）微环境的干预诱导下，其表面干性标志物（如 CD133）的表达会发生显著上调，常被用于研究肺癌干细胞（CSCs）在放化疗抵抗中的分子逃逸机制。

3. 高通量小分子抗癌药物筛选：由于其带有标志性的 TP53 和 PIK3CA 复合突变背景，它是测定多靶点激酶抑制剂、PLK1 抑制剂（如 Rigosertib、Narazaciclib）以及天然植物提取物（如牛至提取物）体外细胞毒性与杀伤增殖动力学（MTT/WST-8 终点检测）的标准肺癌细胞模型之一。

PART 2 ENGLISH SECTION

I General Information and Genetic Background

• Cell Line Name BioVector® NCI-N417

• Synonyms NCI-N-417, NCI-H417, N417, H417, NCIN417, NCI-N417D

• Cellosaurus Accession CVCL_1602

• Species Origin Human Homo sapiens

• Tissue and Disease Background Established by the National Cancer Institute (NCI), this line serves as a classic foundational platform derived from a patient presenting with Small Cell Lung Carcinoma (SCLC). It captures the crucial neuroendocrine features essential for precision bronchopulmonary oncology modeling.

• Genomic and Mutational Profiles

  • Houses a distinct homozygous/hemizygous TP53 nonsense mutation (p.Glu298Ter, introducing an early stop codon).

  • Carries a heterozygous PIK3CA mutation (p.Gln546Lys).

  • Exhibits chromosomal rearrangements leading to a functional PVT1-CLVS1 gene fusion.

  • Contains integrated xenotropic murine leukemia virus-related virus (XMRV) Bxv-1 fragments, reflecting its historical passage through nude mouse xenograft steps.

• Biosafety Level BSL-1 (Standard cell culture precautions; handle according to BSL-2 practices if managing viral transcript tracking).

II Morphological Attributes and Cultivation Media

• Morphology Displays small, high nuclear-to-cytoplasmic ratio profiles typical of the variant subclass of small cell lung cancer cell lines. The biomass possesses almost zero plastic adherence, proliferating synchronously as floating, multicellular suspension clusters and tight spherical aggregates.

• Growth Mode Suspension growth in aggregates.

• Cell Doubling Interval Proliferates dynamically with an approximate doubling period of 26 hours.

• Standard Complete Growth Medium Formulation

  • Basal Medium BioVector® RPMI-1640 medium.

  • Routine Maintenance Supplements 10% premium BioVector® Fetal Bovine Serum. For initial recovery post-thaw, extending parameters to 15% FBS helps protect emerging suspension clusters.

• Physical Incubation Thresholds Regulated strictly at 37 degrees Celsius under an atmospheric layer of 5% Carbon Dioxide and saturated air humidity.

III Subculturing and Thawing Protocols

1. Routine Passaging Schedule

   • Initiate subculturing when the floating aggregate colonies turn dark/dense at their core or when the surrounding medium shows a major pH color shift to yellow.

   • Transfer the cluster suspension into a sterile tube, pellet down at 150 g for 5 minutes, and draw off the exhausted supernatant.

   • Dispense fresh complete BioVector® medium. Disperse large clusters into micro-aggregates by pipetting very gently a few times. Avoid single-cell mechanical shearing, as clustered cell-to-cell signaling is vital for maintaining the line's survival baseline.

   • Seeding Density Maintain viable density between 2.0乘以10的5次方 and 1.0乘以10的6次方 cells/mL. Split ratio ranges normally from 1比2 to 1比4.

2. Cryovial Thawing and Recovery

   • Retrieve the cryovial from liquid nitrogen and submerge it into a 37 degrees Celsius BioVector® water bath with continuous agitation until liquefied within 1 to 2 minutes.

   • Dilute the suspension into a centrifuge tube filled with 5 to 8 mL of pre-warmed complete growth medium and spin down at 150 g for 5 minutes.

   • Decant the DMSO-tainted supernatant, resuspend the pellet in fresh BioVector® complete medium, and seed directly into a fresh culture flask for undisturbed expansion.

IV Strategic Research Applications

1. SCLC Chemoresistance & Phenotypic Shifting: BioVector® NCI-N417 serves as a key reference line to generate cisplatin-resistant clones (e.g., N417/CDDP) and cross-resistance profiles against paclitaxel, driving investigation into ATP-binding cassette (ABC) transporter failures.

2. Cancer Stem Cells (CSCs) & Hypoxic Stress Pathways: Under deliberate hypoxia challenge protocols, this model triggers the up-regulation of the stemness marker CD133, enabling the dissection of hypoxia-inducible factors (HIFs) and drug-evasion mechanisms.

3. High-Throughput Small Molecule Anti-Tumor Screenings: Because it houses concomitant TP53/PIK3CA pathway lesions, it is heavily used to benchmark the cytostatic and anti-proliferative values of checkpoint inhibitors, PLK1 targeting agents (e.g., Rigosertib, Narazaciclib), and novel botanical extracts through standard WST-8 or MTT cytotoxicity matrices.

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