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Show all categories Hep2 Cells and Their Role in Laryngeal Cancer Research Back

Hep2 Cells and Their Role in Laryngeal Cancer Research

Hep 2 cells are a pivotal in vitro model used extensively across biomedical research domains, such as rheumatology, cancer research, and immunology. Originating from laryngeal carcinoma, these human cells have been integral to elucidating the tissue of origin and the specific traits of laryngeal neoplasms. Their significance is well recognized in translational cancer research, where they've greatly contributed to our understanding of the laryngeal nature and origin of cancers, marking a substantial presence in laryngeal cancer research publications [1].

Origin and general characteristics of the Hep 2 cells

A cell line's origin and general characteristics define its applicability in research. This section will help you know about the origin and some salient features of Hep 2 cells. For instance, you will find out: What is the HEp-2 cell line? What is the source of Hep 2 cells? And what is the morphology of Hep 2?

  • Hep 2, an immortal human epithelial cell line was first described by H.W. Toolan as laryngeal carcinoma cells in 1954. However, lately, it has been reported that the Hep 2 cell line is composed of cervical adenocarcinoma cells and originated from contamination of the Hela cell line [2].
  • Hep 2 cells contain Hela marker chromosomes and are found positive for Keratin and human papillomavirus DNA sequences as confirmed via immunoperoxidase staining and PCR, respectively.
  • The Hela cell line derivative Hep 2 possesses an epithelial-like morphology.
  • The Hep 2 cell line exhibits both structural and numerical chromosomal abrasions with a near-triploid karyotype [3].

Division of HeLa cervical cancer cells under microscope.

Hep 2 cell line: Culturing information

Before working with a cell line, we must know the following key points for its culturing. This information can be useful for effectively culturing and maintaining the cell line. You should know: What is the doubling time of HEp-2 cells? Are Hep 2 cells adherent? What is the seeding density of Hep2 cells?

 

Population Doubling Time:

The doubling time reported for Hep 2 cells is approximately 40 hours.

Adherent or in Suspension:

Hep 2 cells are adherent and grow into monolayers.

Seeding Density:

A seeding density of 1 x 104 cells/cm2 is ideal for Hep 2 cell culture. For seeding, adherent Hep 2 cells are rinsed with 1 x PBS solution, followed by incubation with Accutase dissociation solution. After 8–10-minute incubation at ambient temperature, cells are resuspended in media and centrifuged. The collected cells are then dispensed in fresh medium and poured into new flasks for culturing.

Growth Medium:

EMEM or Eagle's minimal essential medium is used to culture Hep 2 cells. This media is supplemented with 10% FBS, 1.0 g/L glucose, 2.2 g/L NaHCO3, 2.0 mM L-glutamine, 1% NEAA, and 1 mM sodium pyruvate for ideal cell growth. Media should be renewed 2 to 3 times a week.

Growth Conditions:

Like other mammalian cell lines, Hep 2 is also cultured in a humidified incubator set at 37°C temperature and with a continuous supply of 5% CO2.

Storage:

Hep 2 cells can be stored in ultra-low temperature electric freezers (below  -150 °C) or in liquid nitrogen vapor phase for long term storage.

Freezing Process and Medium:

The freezing media recommended for Hep 2 cells are CM-1 or CM-ACF. Cells should be frozen using a slow freezing process that allows a gradual 1 °C drop in temperature and protects cell viability.

Thawing Process:

The frozen cells vial is quickly thawed by agitation in water bath at 37°C until a small ice clump is left. Cells are then added to fresh media and centrifuged to remove freezing media components. Later, the cell pellet is resuspended in media, and cells are dispensed into culture flasks. Cells need to rest for almost 24 hours to adhere.

Biosafety Level

Biosafety level 1 laboratory is recommended for handling and maintenance of Hep 2 cell cultures.

Hep 2 cells before and after reaching confluence.

Advantages & Limitations of Hep 2 cells

Almost all cell lines exhibit a unique combination of advantages and limitations that contribute to their use in the research field. This section will describe a few main pros and cons associated with the Hep 2 cell line.

Advantages

The principal advantages of the Hep 2 cell line are:

  • Human Origin: Hep 2 is derived from human epithelial cells, making it a valuable in vitro model for studying human diseases and viral infections.
  • ANA Detection: The Hep 2 cell line possesses a native protein array that presents numerous antigens, making it an excellent substrate for detecting antinuclear antibodies (ANA). This feature enables specific and highly sensitive screening of ANA in serum, making it a crucial diagnostic tool for identifying connective tissue diseases.

Limitations

  • Chromosomal Abnormalities: Hep 2 cells exhibit multiple numerical and structural chromosomal abnormalities. These abnormalities can impact cell behavior and may restrict their applicability in certain laboratory experiments.
  • Tumorigenicity: Hep 2, a tumour-derived human epithelial cell line, may possess genetic abnormalities typically absent in epithelial cells. Consequently, the use of Hep 2 cells might be constrained in specific studies focusing on normal cellular physiology.

Expanding Applications of Hep 2 Cell Line in Biomedical Research

The Hep 2 cell line stands out as an exemplary model for a multitude of applications within biomedical research. Renowned for their versatility, these cells serve critical roles in in vitro experiments, ranging from receptor analysis to the study of complex diseases.

Exploring Tumorigenic Mechanisms and Therapeutic Targets with Hep 2 Cells

Hep 2 cells, being tumorigenic, are pivotal for delving into the intricacies of cancer biology. They provide insights into cancer signaling pathways, mechanistic studies, and are a mainstay in the screening and evaluation of anticancer drugs. For example, an insightful study utilized Hep 2 to delineate the influence of miRNA-33a on cancer cell proliferation. The findings illuminated the antiproliferative effects of miRNA-33a through its interaction with PIM1, a known oncogene, suggesting a novel therapeutic target [4]. In another instance, Hep 2 was employed in assessing the therapeutic potential of Marsdenia tenacissima zinc oxide nanoparticles, highlighting their antiproliferative and apoptotic efficacy [5].

Advancing Virology Research with Hep 2 Cell Insights

The susceptibility of Hep 2 cells to various human viruses makes them an invaluable resource in virological research. They have been effectively used in the expression of SARS-CoV-2 viral genes to unravel the complex interplay between the virus and host cellular mechanisms [6]. This application is particularly crucial in the current era, where understanding and combating viral infections like COVID-19 is a global priority.

Deciphering Cellular Functions: Gene Manipulation in Hep 2 Cells

The Hep 2 cell line's adaptability to genetic manipulation underscores its utility in mechanistic studies. Researchers capitalize on this feature to modulate gene expression and elucidate the roles of specific genes in cellular functions. A notable study involved the overexpression of the RNA-binding protein RBM6 in Hep 2 cells, which facilitated the investigation of its tumor suppressor potential, providing valuable insights into the molecular underpinnings of cancer [7].

Enhancing Disease Diagnosis through Hep 2 Cell Line Applications

Beyond these research domains, Hep 2 cells are acclaimed for their diagnostic capabilities, especially in the detection of ANAs, which are critical in the diagnosis of autoimmune diseases such as systemic lupus erythematosus. The precision with which Hep 2 cells can present ANAs supports the diagnosis and development of targeted treatments, enhancing our understanding of autoimmune pathologies and improving patient care.

Through these diverse applications, Hep 2 cells have significantly contributed to advancements in translational cancer research, the study of viral infections, and the exploration of cellular mechanisms. Their contribution to the generation of clinically relevant data is invaluable, confirming their indispensable role in both the laboratory and the clinic. As research continues to evolve, the Hep 2 cell line is sure to remain at the forefront, aiding in the discovery of new treatments and expanding our knowledge of human health and disease.

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Hep 2 cells: Research Publications

The following are some interesting and most cited research publications on Hep 2 cells.

Resources for Hep2 Cell Line: Protocols, Videos, and More

Hep 2 is a well-known cell line. There are several available resources featuring the Hep 2 cell line.

  • Subculturing Hep2 cell line: This video is a step-by-step guide to subculturing Hep 2 cells.
  • Hep 2 cells ANA screening: This video explains anti-nuclear antibodies (ANA) screening using the Hep 2 cell line.
  • Hep 2 culturing: This link contains basic cell culture information about Hep 2 cells. It includes cell splitting, cell freezing, and thawing.

Frequently Asked Questions about HEp-2 Cells in Biomedical Research

HEp-2 cells are an immortal cell line originally believed to be from laryngeal carcinoma but later identified as cervical adenocarcinoma cells contaminated with HeLa cells. Their human epithelial origin makes them a key tool in studying various diseases, especially in cancer research, immunology, and virology, providing insights into disease mechanisms and potential treatments.
Staining patterns in HEp-2 cells during antinuclear antibody (ANA) testing are critical for diagnosing autoimmune diseases. These patterns help identify specific autoantibodies, aiding in the diagnosis of conditions like systemic lupus erythematosus, and are a cornerstone in clinical immunology for their role in understanding autoimmune pathologies.
HEp-2 cells are widely used for ANA detection due to their ability to present a broad array of antigens, enabling sensitive and specific screening of ANAs in serum samples. This makes them invaluable in diagnosing autoimmune diseases, highlighting their pivotal role in advancing diagnostic methods in immunology.
ANA titres in HEp-2 cells indicate the highest serum dilution at which autoantibodies remain detectable, serving as a crucial measure in assessing autoimmune disease severity and progression. High titres often suggest more active disease, guiding clinicians in treatment decisions and disease monitoring.
Originating from laryngeal carcinoma, HEp-2 cells are vital in exploring laryngeal cancer's origins and traits. Their use in translational cancer research has led to significant advancements in understanding laryngeal cancer, aiding in the development of targeted therapies and enriching laryngeal cancer literature.
Initially thought to be from laryngeal carcinoma, HEp-2 cells were later identified as cervical adenocarcinoma cells, a revelation that highlighted the complexity of cell line origins but did not detract from their value in research due to their unique characteristics and applications in studying epithelial cell behaviors and pathologies.
Despite their common origin from human cervical cancer cells, HeLa and HEp-2 cells display distinct properties. HeLa's rapid proliferation makes it suitable for a broad range of studies, while HEp-2's unique features, particularly in epithelial cell research, offer specific advantages in studying epithelial cell pathologies and mechanisms.
Successful culturing of HEp-2 cells involves specific conditions such as using EMEM supplemented with FBS and maintaining a controlled environment at 37°C with 5% CO2. Knowledge of their culturing requirements, including seeding density and media composition, is essential for their effective use in research.
HEp-2 cells offer numerous advantages, including their relevance in studying human diseases and utility in ANA detection. However, their chromosomal abnormalities and tumorigenic nature can limit their applicability in certain research areas, necessitating careful consideration of their use.
HEp-2 cells have broadened research horizons across various biomedical fields, from drug screening to disease mechanism studies. Their versatility and contributions to understanding and treating diseases underscore their invaluable role in both basic and translational research, promising ongoing relevance in future scientific endeavors.

References

  1. Fusi, M. and S. Dotti, Adaptation of the HEp-2 cell line to totally animal-free culture systems and real-time analysis of cell growth. Biotechniques, 2021. 70(6): p. 319-326.
  2. Gorphe, P., A comprehensive review of Hep-2 cell line in translational research for laryngeal cancer. Am J Cancer Res, 2019. 9(4): p. 644-649.
  3. Wang, M., et al., Cancer-associated fibroblasts in a human HEp-2 established laryngeal xenografted tumor are not derived from cancer cells through epithelial-mesenchymal transition, phenotypically activated but karyotypically normal. PLoS One, 2015. 10(2): p. e0117405.
  4. Karatas, O.F., Antiproliferative potential of miR-33a in laryngeal cancer Hep-2 cells via targeting PIM1. Head Neck, 2018. 40(11): p. 2455-2461.
  5. Wang, Y., et al., Synthesis of Zinc oxide nanoparticles from Marsdenia tenacissima inhibits the cell proliferation and induces apoptosis in laryngeal cancer cells (Hep-2). Journal of Photochemistry and Photobiology B: Biology, 2019. 201: p. 111624.
  6. Zhang, J., et al., A systemic and molecular study of subcellular localization of SARS-CoV-2 proteins. Signal Transduct Target Ther, 2020. 5(1): p. 269.
  7. Wang, Q., et al., RNA-binding protein RBM6 as a tumor suppressor gene represses the growth and progression in laryngocarcinoma. Gene, 2019. 697: p. 26-34.
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