BioVector? NCI-N417 人小細(xì)胞肺癌細(xì)胞株

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

第一部分 中文說明

一 產(chǎn)品基本信息與遺傳學(xué)背景

• 細(xì)胞名稱 BioVector? NCI-N417 人小細(xì)胞肺癌細(xì)胞

• 別名與同義詞 NCI-N-417、NCI-H417、N417、H417、NCIN417

• 系統(tǒng)學(xué) Accession Cellosaurus CVCL_1602

• 物種來源 人類 Homo sapiens

• 組織與疾病背景 該細(xì)胞株由美國國家癌癥研究所（NCI）的 Gazdar 及其合作者建立，源自一名確診為肺小細(xì)胞癌（SCLC）患者的病灶，是研究經(jīng)典神經(jīng)內(nèi)分泌型肺癌的標(biāo)志性體外模型之一。

• 突變特征與基因組狀態(tài)

  • 帶有明確的 TP53 基因純合/半合子突變（p.Glu298Ter，導(dǎo)致蛋白翻譯提前終止）。

  • 帶有 PIK3CA 基因雜合突變（p.Gln546Lys）。

  • 染色體易位 包含 PVT1-CLVS1 基因融合。

  • 特殊特征 包含集成的異種小鼠白血病病毒相關(guān)病毒（XMRV）Bxv-1，提示其在早期建立過程中可能經(jīng)過了裸鼠異種移植階段。

• 生物安全級別 1級（或根據(jù)其攜帶的內(nèi)源反轉(zhuǎn)錄病毒片段成分，遵循常規(guī) 2級生物安全防護(hù)規(guī)范操作更為穩(wěn)妥）。

二 細(xì)胞形態(tài)學(xué)與培養(yǎng)環(huán)境

• 形態(tài)學(xué)特征 展現(xiàn)典型的小細(xì)胞肺癌變異亞型（Variant subclass）形態(tài)特征。細(xì)胞體積相對較小，核質(zhì)比極高。它們在培養(yǎng)基中極少貼壁，主要以松散的懸浮細(xì)胞團(tuán)塊、密集聚集體形式在液體中漂浮生長。

• 生長模式 懸浮聚集生長。

• 倍增時(shí)間 增殖較快，倍增時(shí)間大約為 26小時(shí)。

• 標(biāo)準(zhǔn)完全培養(yǎng)基配方

  • 基礎(chǔ)培養(yǎng)基 BioVector? RPMI-1640 培養(yǎng)基。

  • 維持添加 10% BioVector? 優(yōu)質(zhì)胎牛血清（對于狀態(tài)不佳或新復(fù)蘇的細(xì)胞，可選用 15% 胎牛血清以加速恢復(fù)）。

• 物理培養(yǎng)參數(shù) 37攝氏度恒溫、5% 二氧化碳、空氣飽和濕度。

三 細(xì)胞傳代與復(fù)蘇標(biāo)準(zhǔn)操作步驟

1. 常規(guī)傳代操作

   • 當(dāng)懸浮的細(xì)胞簇生長得過于龐大、內(nèi)部開始出現(xiàn)發(fā)黑的多重細(xì)胞重疊壞死，或者培養(yǎng)基顯著變黃時(shí)需要進(jìn)行傳代。

   • 將包含細(xì)胞團(tuán)的懸液轉(zhuǎn)移至離心管中，以 150 g 離心 5分鐘，棄去舊培養(yǎng)基。

   • 用新鮮的 BioVector? 完全培養(yǎng)基重懸細(xì)胞。傳代時(shí)必須使用移液槍非常輕柔地吹打幾次，將過大的團(tuán)塊打散成大小適中的小細(xì)胞叢。切勿使用機(jī)械手段強(qiáng)行徹底打散成絕對的單細(xì)胞，否則會(huì)破壞細(xì)胞間的集聚信號導(dǎo)致大批細(xì)胞死亡。

   • 推薦接種密度 維持在每毫升 2.0乘以10的5次方 至 1.0乘以10的6次方 個(gè)活細(xì)胞。傳代比例通常為 1比2 至 1比4。

2. 凍存細(xì)胞復(fù)蘇

   • 從液氮中取出冷凍管，立即投入 37攝氏度 BioVector? 水浴鍋中快速搖動(dòng)使其融化，控制在 1到2分鐘內(nèi)。

   • 將解凍的細(xì)胞懸液移至含 5到8 mL 預(yù)熱培養(yǎng)基的離心管中，以 150 g 離心 5分鐘。

   • 棄去含有二甲基亞砜（DMSO）的舊上清，加入新鮮 BioVector? 完全培養(yǎng)基重懸，接種到培養(yǎng)瓶內(nèi)，靜置培養(yǎng)。

四 核心科研應(yīng)用方向

1. 小細(xì)胞肺癌（SCLC）耐藥株的創(chuàng)制與逆轉(zhuǎn)研究：BioVector? NCI-N417 廣泛用于構(gòu)建抗順鉑（Cisplatin-resistant, 如 N417/CDDP 細(xì)胞）和抗紫杉醇（Paclitaxel）的耐藥模型，用于篩選多藥耐藥相關(guān)蛋白（MRP）表達(dá)和拓?fù)洚悩?gòu)酶突變引起的靶向脫靶現(xiàn)象。

2. 腫瘤干細(xì)胞與缺氧微環(huán)境研究：該細(xì)胞在缺氧（Hypoxia）微環(huán)境的干預(yù)誘導(dǎo)下，其表面干性標(biāo)志物（如 CD133）的表達(dá)會(huì)發(fā)生顯著上調(diào)，常被用于研究肺癌干細(xì)胞（CSCs）在放化療抵抗中的分子逃逸機(jī)制。

3. 高通量小分子抗癌藥物篩選：由于其帶有標(biāo)志性的 TP53 和 PIK3CA 復(fù)合突變背景，它是測定多靶點(diǎn)激酶抑制劑、PLK1 抑制劑（如 Rigosertib、Narazaciclib）以及天然植物提取物（如牛至提取物）體外細(xì)胞毒性與殺傷增殖動(dòng)力學(xué)（MTT/WST-8 終點(diǎn)檢測）的標(biāo)準(zhǔn)肺癌細(xì)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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