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NCI-H460 Cells - Navigating Lung Cancer Research with NCI-H460 Insights

NCI-H460 are human-derived non-small cell lung cancer cells commonly used in lung carcinoma and toxicology research. This cell line is a valuable tool for studying various cancer biology aspects involving tumour development, growth, and drug resistance. Moreover, NCI-H460 cells are an appropriate model for developing anti-cancer reagents.

Before working with the NCI-H460 cell line, this article will help you learn some basic and advanced concepts related to it. Mainly, you will go through:

  1. NCI-H460 cells: Origin and general characteristics
  2. NCI-H460 cell line culturing information
  3. Advantages & disadvantages of NCI-H460 cell line
  4. Research applications of NCI-H460 cells
  5. Research Publications Featuring NCI-H460 Cells
  6. Resources for NCI-H460 Cell line: Protocols, Videos, and More

1.      NCI-H460 cells: Origin and general characteristics

The origin and general characteristics of a cell line majorly contribute to its research applications. This section of the article will assist you in learning the origin and salient features of NCI-H460 lung cancer cells. You will know: What are NCI-H460 cells? What is the NCI-H460 cell line type? What is the morphology of NCI-H460?

  • The NCI-H460 cell line originated from the pleural effusion of a European male with large-cell lung cancer. It was established in 1982 by A.F. Gazdar and colleagues.
  • NCI-H460 lung cancer cells possess an epithelial morphology.
  • NCI-H460 is a tumorigenic cell line with a hypotriploid karyotype. The modal chromosome number for these cells is 57. NCI-H460 cells also possess 58 modal chromosome numbers at comparable rates.
  • These lung cancer cells have many NCI-NCI-H460 mutations like non-small cell lung tumours, such as NCI-H460 KRAS mutation, which are involved in cell proliferation, growth, invasion, and metastasis.

A549 Vs NCI-H460

Both are non-small lung cancer cell lines. They also have similar mutations. The main difference between these cell lines is: NCI-H460 possesses a high growth rate (double) than the A549 cell line.

The CT scan of the middle-aged patient showed a lung tumor in the left lower lobe of the lung. After further examinations, small cell lung carcinoma (SCLC) was diagnosed.

2.      NCI-H460 Cells Culturing Information

You should know the following key points for proper handling and maintenance of the NCI-H460 cell line. It will inform you about NCI-H460 doubling time, NCI-H460 culture medium, and basic cell culture procedures for NCI-H460 lung cancer cells.

Key Points for Culturing NCI-H460 Cells

Doubling Time:

The NCI-H460 doubling time is approximately 33 hours.

Adherent or in Suspension:

The NCI-H460 lung cancer cells are adherent.

Sub-cultivation Ratio:

The split ratio recommended for the NCI-H460 cell line is 1:2 and 1:4. After removing the old medium, adherent cells are rinsed with 1 x phosphate buffer saline. Afterwards, cells are incubated with Accutase passaging solution for 8 to 10 minutes at ambient temperature. Dissociated cells are resuspended in a culture medium and centrifuged. Harvested cells are again resuspended and poured into the new flask for culturing.

Growth Medium:

RPMI 1640 is used as NCI-H460 culture medium. It is supplemented with 10% fetal bovine serum, 2.1 mM stable Glutamine, and 2.0 g/L NaHCO3. Media should be replaced 2 to 3 times a week.

Growth Conditions:

NCI-H460 cultures are maintained at 37°C temperature in a humidified incubator with a continuous 5% CO2 supply.


NCI-H460 lung cancer cells can be stored in the vapour phase of liquid nitrogen or at below -150°C temperature in an electric ultra-low temperature freezer for the long term.

Freezing Process and Medium:

CM-1 or CM-ACF medium is used to freeze and store NCI-H460 cells. A slow freezing method is recommended to protect maximum cell viability.

Thawing Process:

Frozen NCI-H460 cells are thawed in a pre-warmed water bath (at 37°C temperature) for 40 to 60 seconds until a small ice clump is left. Thawed cells are added with fresh media and centrifuged to remove freezing media components. The collected cell pellet is again resuspended, and cells are dispensed into new flasks containing media for growth. NCI-H460 cells may take almost 24 hours to attach to the surface of the flask.

Biosafety Level:

NCI-H460 lung cancer cells are handled and maintained in biosafety level 1 laboratories.


NCI-H460 cells growing together in adherent clusters at 10x and 20x magnification.

3.      Advantages & Disadvantages of NCI-H460 cells

NCI-H460 is a widely used cell line in lung cancer research. This section will cover the general advantages and disadvantages related to NCI-H460 lung cancer cells.


The advantages of the NCI-H460 non-small cell lung cancer cell line are:

Tumour origin

The NCI-H460 cell line was derived from a large-cell lung carcinoma patient, representing this particular lung cancer type. They are used as a model to study lung cancer biology and develop new and effective treatments. NCI-H460 cells have tumorigenic potential and can be injected in immunodeficient mice to create in vivo tumour models for studying tumour growth, development, and efficacy of potential drugs.

High proliferation rates

NCI-H460 exhibits higher growth rates than other non-small cell lung cancer cell lines, such as A549. This advantage increases their availability and helps researchers conduct reproducible and time-sensitive experiments.



The disadvantages associated with NCI-H460 lung cancer cells are:


NCI-H460 cells are homogenous as they were obtained from a single patient tumour. Therefore, they generally lack complexity and heterogeneity observed in patient tumours.


4.      Research applications of NCI-H460 cells

The NCI-H460 lung cancer cells are extensively used in studies related to lung carcinoma. Here are some of the important research applications of NCI-H460 cells:

  • Lung cancer research: NCI-H460 cells are an invaluable model for investigating cell and molecular mechanisms involved in tumour development, growth, and metastasis. Moreover, it is used to study imperative signalling pathways, molecular targets, and various genetic mutations associated with lung cancer progression. Several studies have been conducted on NCI-H460 cells to study these factors effectively. A study conducted in 2019 proposed that overexpressed nuclear ubiquitous casein and cyclin-dependent kinases substrate (NUCKS) are involved in tumour cell growth via regulation of the PI3K/AKT signalling pathway [1]. Similarly, an in vitro and in vivo study employed NCI-H460 cells to study the role of the eIF4E gene. The findings revealed that the eIF4E gene is involved in lung carcinoma growth and angiogenesis, and it can be targeted to develop promising anti-lung cancer drugs [2].
  • Drug discovery and development: NCI-H460, a human lung cancer cell line, is widely used in drug discovery and development studies. Researchers use these cells to investigate the toxicity and efficacy of novel drug candidates, targeted therapies, and treatments mainly targeting NCI-H460 KRAS mutations. A study conducted by Haoyue Hu and colleagues in 2023 employed NCI-H460 cells to study the anti-cancer effects of the anlotinib drug. The results showed that anlotinib partly affected the growth of KRAS mutant lung cancer cells by inhibiting the MEK/ERK signalling cascade [3]. Likewise, a phenolic compound, carnosic acid was screened for antiproliferative and proapoptotic activities using NCI-H460 cells [4].
  • Drug resistance: The NCI-H460 cell line is ideal for studying lung carcinoma's drug resistance mechanism. Researchers use these cells to develop drug resistance models to identify underlying genes, molecular factors, and signalling pathways. Such as, a study developed Pemetrexed, a multitargeted antifolate agent, resistant NCI-H460 cells to study underlying molecular mechanisms of pemetrexed resistance in non-small cell lung cancer cells [5].

5.      Research Publications Featuring NCI-H460 Cells

Here are some interesting research publications about the NCI-H460 lung carcinoma cell line.

A Natural Glucan from Black Bean Inhibits Cancer Cell Proliferation via PI3K-Akt and MAPK Pathway

This study in the Molecules (2023) proposed that a natural α-1,6-glucan, BBWPW, from black bean inhibits the proliferation of NCI-H460 cells via regulating PI3K/AKT/MAPK pathway.

Dioscin-6′-O-acetate inhibits lung cancer cell proliferation via inducing cell cycle arrest and caspase-dependent apoptosis

This article in Phytomedicine (2019) studied that dioscin-6′-O-acetate, a novel natural compound, exerts an antiproliferative effect on NCI-H460 lung cancer cells.

miRNA-425-5p enhances lung cancer growth via the PTEN/PI3K/AKT signaling axis

BMC Pulmonary Medicine (2020) research states that microRNA-425-5p enhances lung carcinoma tumorigenesis through PTEN/PI3K/AKT pathway.

Quinalizarin exerts an anti-tumour effect on lung cancer A549 cells by modulating the Akt, MAPK, STAT3 and p53 signalling pathways

This article in Molecular Medicine Reports (2017) proposed the antitumor potential and underlying mechanisms of the quinalizarin compound in NCI-H460 and other lung cancer cells.

Eucalyptus globulus Labill. decoction extract inhibits the growth of NCI-H460 cells by increasing the p53 levels and altering the cell cycle profile

This research in the Food and Function (2019) highlights the potential anti-cancer effect of Eucalyptus globulus labill. extract using NCI-H460 cells. The findings showed that plant extract showed these effects by increasing NCI-H460 p53 expression and modulating the cell cycle profile.

6.      Resources for NCI-H460 Cell line: Protocols, Videos, and More

Here are some online resources featuring the NCI-H460 lung cancer cells.

The following links contain imperative cell culture information for H460 cells. 

  • NCI-H460 cells: This website will provide important information about NCI-H460 cell media, subculturing, freezing, and thawing procedures.
  • Passaging NCI-H460 cells: This document will guide you about the passaging and subculturing of the NCI-H460 cell line. Besides, it will also help you learn transfection protocol for NCI-H460 cells.


  1. Hu, C., et al., NUCKS Promotes the Proliferation, Migration and Invasion of Lung Cancer Cells Through Pi3k/Akt Signalling Pathway. Clinical and Investigative Medicine, 2021. 44(2): p. E55-61.
  2. Qi, X., et al., EGPI-1, a novel eIF4E/eIF4G interaction inhibitor, inhibits lung cancer cell growth and angiogenesis through Ras/MNK/ERK/eIF4E signaling pathway. Chemico-Biological Interactions, 2022. 352: p. 109773.
  3. Hu, H., et al., Anlotinib exerts anti-cancer effects on KRAS-mutated lung cancer cell through suppressing the MEK/ERK pathway. Cancer Management and Research, 2020: p. 3579-3587.
  4. Corveloni, A.C., et al., Carnosic acid exhibits antiproliferative and proapoptotic effects in tumoral NCI-H460 and nontumoral IMR-90 lung cells. Journal of Toxicology and Environmental Health, Part A, 2020. 83(10): p. 412-421.
  5. Xu, Y.-L., et al., Establishment and characterization of pemetrexed-resistant NCI-H460/PMT cells. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents), 2019. 19(6): p. 731-739.


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