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NCI-H69AR
Tissue
轉移性
Disease
肺小細胞癌
Metastatic site
胸腔積液
Synonyms
NCI-H69 AR, NCI-H69/AR, H69AR, H-69AR

Documents

NCI-H69AR

Cell line · Product number 305840

NCI-H69AR is a multidrug-resistant derivative of the parental small cell lung carcinoma (SCLC) cell line NCI-H69. It was developed through continuous sele...

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  • CoA
1

01General information

Description

NCI-H69AR 是源自小細胞肺癌(SCLC)親本細胞系 NCI-H69 的多重抗藥性衍生物。它是透過在漸增濃度的化療藥物(如多柔比星)中進行持續篩選而開發出來的。 因此,NCI-H69AR 成為研究小細胞肺癌(SCLC)後天藥物抗藥性機制的重要模型系統。該細胞系保留了其親代細胞系的許多形態學和生化特徵,但對多種細胞毒性藥物表現出顯著的抗藥性,使其特別適用於研究外排介導的抗藥性途徑。

NCI-H69AR 的主要抗藥機制涉及由 MDR1 基因編碼的多藥抗性蛋白 P-糖蛋白(P-gp)的過度表達。 P-gp 作為一種 ATP 依賴性外排泵,能降低細胞內藥物的累積,特別是對蒽環類、長春鹼類及附生黴素類藥物。 此外,NCI-H69AR 展現出膜相關蛋白質(包括 annexin II)的表達改變,這可能與鈣訊號傳導及囊泡運輸的變化有關——這些過程皆與藥物抗藥性及細胞應激反應有關。 這些表型改變使 NCI-H69AR 成為識別藥物抗性調控因子,以及評估針對外排機制或完全繞過抗性路徑之藥物療效的寶貴模型。

NCI-H69AR 亦曾與其親本系進行比較研究,用以闡明基因與蛋白質表達、藥物敏感性特徵,以及對藥理學抑制劑反應的變化。此比較框架有助於釐清癌症中藥物抗藥性的演變,並有助於設計旨在使抗藥性腫瘤恢復敏感性的聯合療法。 該細胞株通常在添加胎牛血清的 RPMI-1640 培養基中培養,並在標準大氣條件下維持。其穩健的特性與特徵明確的抗藥性表型,使其在肺癌抗藥性的臨床前研究中佔有一席之地。

Tissue
轉移性
Disease
肺小細胞癌
Metastatic site
胸腔積液
Synonyms
  • NCI-H69 AR
  • NCI-H69/AR
  • H69AR
  • H-69AR

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮性
Cell type
類上皮性
Tumourigenic
是的; 是的,在裸鼠身上

04Identifiers

Citation
NCI-H69AR(Cytion 型號 305840)
NCBI TaxID
9606
Cellosaurus
CVCL_3513

05Expression and susceptibility

Mutational profile
突變:PIK3CA,簡單突變,p.Gly106_Arg108del (c.317_325delGGCAACCGT),雜合子(源自親代細胞系)。突變,RB1,簡單型,p.Glu748Ter (c.2242G>T),純合子(源自親本細胞系)。突變,TP53,簡單型,p.Glu171Ter (c.511G>T),純合子(源自親代細胞系)。

06Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 20% 的胎牛血清 (FBS)
Dissociation reagent
Accutase
Fluid renewal
每週 2 至 3 次
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

08Donor

Donor age
55年
Ethnicity
高加索人