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11/04/2023

LNCaP cells

LNCaP (Lymph Node Carcinoma of the Prostate) is a commonly used human cell line in the cancer research field. These cells are extensively applied to study the biology of prostate cancer and to evaluate potential treatments.

This article will assist you to go through all the basic information about the LNCaP cell line, crucial for you to start working with it. You will learn:

  1. Characteristics and origin of LNCaP cells
  2. Advantages & Disadvantages of LNCaP
  3. Research Applications of LNCaP cells
  4. Resources for LNCaP cells
  5. Characteristics and origin of LNCaP cells

In this section, we will talk about the origin and distinguishing characteristics of the LNCaP cell line.

  • LNCaP cells are an androgen-sensitive cell line that was first isolated from a patient with prostate cancer in 1977.
  • The cells were derived from the left supraclavicular metastatic lymph node of a 50-year-old Caucasian male [1].
  • LNCaP cells form subcutaneous tumors in male athymic nude mice with a frequency of 58%.
  • The cells are epithelial-like in shape and can grow in aggregates or as single cells.
  • The average size of LNCaP prostate cancer cells is 18 µm.
  • LNCaP cells are aneuploid, with a modal number of chromosomes ranging between 76 to 91.
  • Two sublines have been derived from the original LNCaP cells: LNCaP-G4 (high androgen sensitive) and LNCaP-E9 (low androgen sensitive), which are suitable for prostate cancer studies [2].

Prostate cells in SEM

LNCaP culturing information

Before starting with these prostate cancer cells, you need to keep in mind the following LNCaP cell culture basics.

Specifically, you should know: What is the doubling time of LNCaP cells? What media to use for LNCaP cells? What is the seeding density of LNCaP? 

Key Points for Culturing LNCaP Cells

Population Doubling Time: The average population doubling time for LNCaP is 48 to 60 hours.

Adherent or in Suspension: LNCaP are adherent, grow in monolayers and form cell clusters.

Seeding Density: The recommended seeding density is 1-2 x 104 cells/cm2. The adherent LNCaP cells are washed with PBS and detached by Accutase. Next, centrifuge, resuspend, and culture in new flasks containing growth media.

Growth Medium: EMEM media supplemented with 2.5 mM L-glutamine and 10% fetal bovine serum (FBS) is used to culture LNCaP cells. Media should be renewed every 3rd day.  

Growth Conditions (Temperature, CO2): LNCaP cells optimally grow in a humidified incubator set at 37°C temperature and with a 5% CO2 supply.

Storage: Keep in the vapour phase of liquid nitrogen to maintain cell viability.

Freezing Process and Medium: CM-1 or CM-ACF freeze medium is used to freeze LNCaP cells. A slow freezing process is recommended to protect cells from shock.

Thawing Process: Thaw in a 37°C water bath until a small ice clump remains. Resuspend cells in growth medium, centrifuge to remove freeze medium and resuspend with growth medium and transfer into the flask for culturing.

Biosafety Level: Biosafety level 1 is recommended to maintain the LNCaP cell culture.

LNCaP Cells at low and high confluency

2.       Advantages & Disadvantages of LNCaP

LNCaP is an extensively used prostate cancer cell line. Advantages and disadvantages of this cell line are discussed here.

Advantages

  • Easy to culture: LNCaP cells are easy to culture and maintain in a research laboratory, making them broadly used in drug screening and testing purposes.
  • Androgen sensitivity: LNCaP is an androgen-dependent cell line that expresses androgen receptors (AR) and requires androgen hormones to grow and proliferate. They are commonly used as an in vitro model to study androgen-dependent prostate cancer.

Disadvantages

  • Slow growth: LNCaP cells have a slow growth rate, making them sometimes unsuitable for some experimental settings.
  • Cell clusters: LNCaP cells mostly form cell aggregates and clusters that can affect readout in several cell-based assays.

3.       Research Applications of LNCaP cells

Some of the important research applications of LNCaP cells are:

  1. Prostate cancer research: LNCaP represents an androgen-dependent cell model of prostate cancer. Thus, it is used to study androgen receptors and other gene pathway signaling in prostate cancer. A study used CRISPR/Cas9 to cleave the AR gene in LNCaP cells and observed its effect on prostate cancer cell growth. The findings suggest that targeting AR can suppress the growth of androgen-sensitive cancer cells [3]. Another study found that six transmembrane epithelial antigen of the prostate 1 (STEAP1) gene is mostly unregulated in prostate cancer cells and is responsible for cancer cell growth. Silencing of the STEAP1 gene inhibits LNCaP cell growth and induces apoptosis by counteracting the effect of the androgens [4].
  2. Drug discovery: The LNCaP cell line is extensively used to screen anti-cancer drugs. Several studies have used this prostate cancer model to evaluate the therapeutic effects of potential drug candidates. Tousi and colleagues examined the apoptotic effect of Eupatorin containing mPEG-b-PLGA coated iron oxide nanoparticles on LNCaP prostate cancer cells. The results revealed that this form of Eupatorin is more effective in inhibiting cancer cell growth than the free form [5].

4.       Resources for LNCaP cells

Prostate cells under microscope

Research Publications with LNCaP cells as a research model

There is a plethora of research studies on LNCaP cells. Some interesting research publications are mentioned here.

CRISPR/Cas9 targeting of the androgen receptor suppresses the growth of LNCaP human prostate cancer cells: This paper was published in Molecular medicine reports in 2018. This study used CRISPR/Cas9 as a therapeutic tool to target androgen receptor (AR) and inhibit the growth of LNCaP prostate cancer cells.

Phytochemical analysis of Daphne pontica L. stems with their pro-apoptotic properties against DU-145 and LNCaP prostate cancer cells: This publication in the DARU Journal of Pharmaceutical Sciences proposed that Daphne pontica L. plant stems possess anti-cancer potential against LNCaP and DU-145 prostate cancer cells.

Synergistic anti-tumor effects of Liraglutide, a glucagon-like peptide-1 receptor agonist, along with Docetaxel on LNCaP prostate cancer cell line: This study was published in the European Journal of Pharmacology in 2020. This research proposed that glucagon-like peptide-1 receptor agonist, Liraglutide, and Docetaxel drug synergism can cause a potent anti-tumor effect on LNCaP cells.

The determination of the potential anticancer effects of Coriandrum sativum in PC‐3 and LNCaP prostate cancer cell lines: This paper in the Journal of Cellular Biochemistry (2018) proposed that Coriandrum sativum extract exerts an anticancer effect on LNCaP cells. It modulates the expression of certain cell cycle genes to regulate cell proliferation, migration, and invasion.

Knockdown of STEAP1 inhibits cell growth and induces apoptosis in LNCaP prostate cancer cells counteracting the effect of androgens: This research article was published in the Molecular oncology journal in 2018. This study explored that STEAP1 is overexpressed in most cancer cells including LNCaP cells and regulates cancer cell growth. Inhibition or knockdown of this gene can prevent the growth of prostate cancer cells and also induce cell death. 

LNCaP cells: Protocols, Videos, and More

LNCaP is a commonly used prostate cancer cell line with many resources available on this cell line including extensive culturing and transfection protocols.

Transfecting LNCaP cell line: This link will help you learn LNCaP cell passaging and transfection protocol.

LNCaP cell transfection: This website contains a protocol for LNCaP cell transfection with plasmid DNA.

Transfection tutorial LNCaP: This video is a step-by-step guide for learning the transfection protocol of the LNCaP cell line.

Culturing LNCaP cells: This document provides a protocol for subculturing the LNCaP prostate cancer cell line.

References

  1. Castanares, M.A., et al., Characterization of a novel metastatic prostate cancer cell line of LNCaP origin. The Prostate, 2016. 76(2): p. 215-225.
  2. Iguchi, K., et al., Isolation and characterization of LNCaP sublines differing in hormone sensitivity. Journal of andrology, 2007. 28(5): p. 670-678.
  3. Wei, C., et al., CRISPR/Cas9 targeting of the androgen receptor suppresses the growth of LNCaP human prostate cancer cells. Molecular medicine reports, 2018. 17(2): p. 2901-2906.
  4. Gomes, I.M., et al., Knockdown of STEAP1 inhibits cell growth and induces apoptosis in LNCaP prostate cancer cells counteracting the effect of androgens. Medical Oncology, 2018. 35: p. 1-10.
  5. Tousi, M.S., et al., Evaluation of apoptotic effects of mPEG-b-PLGA coated iron oxide nanoparticles as a eupatorin carrier on DU-145 and LNCaP human prostate cancer cell lines. Journal of Pharmaceutical Analysis, 2021. 11(1): p. 108-121.

 

LNCaP (Lymph Node Carcinoma of the Prostate) is a commonly used human cell line in the cancer research field. These cells are extensively applied to study the biology of prostate cancer and to evaluate potential treatments.

This article will assist you to go through all the basic information about the LNCaP cell line, crucial for you to start working with it. You will learn:

  1. Characteristics and origin of LNCaP cells
  2. Advantages & Disadvantages of LNCaP
  3. Research Applications of LNCaP cells
  4. Resources for LNCaP cells
  5. Characteristics and origin of LNCaP cells

In this section, we will talk about the origin and distinguishing characteristics of the LNCaP cell line.

  • LNCaP cells are an androgen-sensitive cell line that was first isolated from a patient with prostate cancer in 1977.
  • The cells were derived from the left supraclavicular metastatic lymph node of a 50-year-old Caucasian male [1].
  • LNCaP cells form subcutaneous tumors in male athymic nude mice with a frequency of 58%.
  • The cells are epithelial-like in shape and can grow in aggregates or as single cells.
  • The average size of LNCaP prostate cancer cells is 18 µm.
  • LNCaP cells are aneuploid, with a modal number of chromosomes ranging between 76 to 91.
  • Two sublines have been derived from the original LNCaP cells: LNCaP-G4 (high androgen sensitive) and LNCaP-E9 (low androgen sensitive), which are suitable for prostate cancer studies [2].

Prostate cells in SEM

LNCaP culturing information

Before starting with these prostate cancer cells, you need to keep in mind the following LNCaP cell culture basics.

Specifically, you should know: What is the doubling time of LNCaP cells? What media to use for LNCaP cells? What is the seeding density of LNCaP? 

Key Points for Culturing LNCaP Cells

Population Doubling Time: The average population doubling time for LNCaP is 48 to 60 hours.

Adherent or in Suspension: LNCaP are adherent, grow in monolayers and form cell clusters.

Seeding Density: The recommended seeding density is 1-2 x 104 cells/cm2. The adherent LNCaP cells are washed with PBS and detached by Accutase. Next, centrifuge, resuspend, and culture in new flasks containing growth media.

Growth Medium: EMEM media supplemented with 2.5 mM L-glutamine and 10% fetal bovine serum (FBS) is used to culture LNCaP cells. Media should be renewed every 3rd day.  

Growth Conditions (Temperature, CO2): LNCaP cells optimally grow in a humidified incubator set at 37°C temperature and with a 5% CO2 supply.

Storage: Keep in the vapour phase of liquid nitrogen to maintain cell viability.

Freezing Process and Medium: CM-1 or CM-ACF freeze medium is used to freeze LNCaP cells. A slow freezing process is recommended to protect cells from shock.

Thawing Process: Thaw in a 37°C water bath until a small ice clump remains. Resuspend cells in growth medium, centrifuge to remove freeze medium and resuspend with growth medium and transfer into the flask for culturing.

Biosafety Level: Biosafety level 1 is recommended to maintain the LNCaP cell culture.

LNCaP Cells at low and high confluency

2.       Advantages & Disadvantages of LNCaP

LNCaP is an extensively used prostate cancer cell line. Advantages and disadvantages of this cell line are discussed here.

Advantages

  • Easy to culture: LNCaP cells are easy to culture and maintain in a research laboratory, making them broadly used in drug screening and testing purposes.
  • Androgen sensitivity: LNCaP is an androgen-dependent cell line that expresses androgen receptors (AR) and requires androgen hormones to grow and proliferate. They are commonly used as an in vitro model to study androgen-dependent prostate cancer.

Disadvantages

  • Slow growth: LNCaP cells have a slow growth rate, making them sometimes unsuitable for some experimental settings.
  • Cell clusters: LNCaP cells mostly form cell aggregates and clusters that can affect readout in several cell-based assays.

3.       Research Applications of LNCaP cells

Some of the important research applications of LNCaP cells are:

  1. Prostate cancer research: LNCaP represents an androgen-dependent cell model of prostate cancer. Thus, it is used to study androgen receptors and other gene pathway signaling in prostate cancer. A study used CRISPR/Cas9 to cleave the AR gene in LNCaP cells and observed its effect on prostate cancer cell growth. The findings suggest that targeting AR can suppress the growth of androgen-sensitive cancer cells [3]. Another study found that six transmembrane epithelial antigen of the prostate 1 (STEAP1) gene is mostly unregulated in prostate cancer cells and is responsible for cancer cell growth. Silencing of the STEAP1 gene inhibits LNCaP cell growth and induces apoptosis by counteracting the effect of the androgens [4].
  2. Drug discovery: The LNCaP cell line is extensively used to screen anti-cancer drugs. Several studies have used this prostate cancer model to evaluate the therapeutic effects of potential drug candidates. Tousi and colleagues examined the apoptotic effect of Eupatorin containing mPEG-b-PLGA coated iron oxide nanoparticles on LNCaP prostate cancer cells. The results revealed that this form of Eupatorin is more effective in inhibiting cancer cell growth than the free form [5].

4.       Resources for LNCaP cells

Prostate cells under microscope

Research Publications with LNCaP cells as a research model

There is a plethora of research studies on LNCaP cells. Some interesting research publications are mentioned here.

CRISPR/Cas9 targeting of the androgen receptor suppresses the growth of LNCaP human prostate cancer cells: This paper was published in Molecular medicine reports in 2018. This study used CRISPR/Cas9 as a therapeutic tool to target androgen receptor (AR) and inhibit the growth of LNCaP prostate cancer cells.

Phytochemical analysis of Daphne pontica L. stems with their pro-apoptotic properties against DU-145 and LNCaP prostate cancer cells: This publication in the DARU Journal of Pharmaceutical Sciences proposed that Daphne pontica L. plant stems possess anti-cancer potential against LNCaP and DU-145 prostate cancer cells.

Synergistic anti-tumor effects of Liraglutide, a glucagon-like peptide-1 receptor agonist, along with Docetaxel on LNCaP prostate cancer cell line: This study was published in the European Journal of Pharmacology in 2020. This research proposed that glucagon-like peptide-1 receptor agonist, Liraglutide, and Docetaxel drug synergism can cause a potent anti-tumor effect on LNCaP cells.

The determination of the potential anticancer effects of Coriandrum sativum in PC‐3 and LNCaP prostate cancer cell lines: This paper in the Journal of Cellular Biochemistry (2018) proposed that Coriandrum sativum extract exerts an anticancer effect on LNCaP cells. It modulates the expression of certain cell cycle genes to regulate cell proliferation, migration, and invasion.

Knockdown of STEAP1 inhibits cell growth and induces apoptosis in LNCaP prostate cancer cells counteracting the effect of androgens: This research article was published in the Molecular oncology journal in 2018. This study explored that STEAP1 is overexpressed in most cancer cells including LNCaP cells and regulates cancer cell growth. Inhibition or knockdown of this gene can prevent the growth of prostate cancer cells and also induce cell death. 

LNCaP cells: Protocols, Videos, and More

LNCaP is a commonly used prostate cancer cell line with many resources available on this cell line including extensive culturing and transfection protocols.

Transfecting LNCaP cell line: This link will help you learn LNCaP cell passaging and transfection protocol.

LNCaP cell transfection: This website contains a protocol for LNCaP cell transfection with plasmid DNA.

Transfection tutorial LNCaP: This video is a step-by-step guide for learning the transfection protocol of the LNCaP cell line.

Culturing LNCaP cells: This document provides a protocol for subculturing the LNCaP prostate cancer cell line.

References

  1. Castanares, M.A., et al., Characterization of a novel metastatic prostate cancer cell line of LNCaP origin. The Prostate, 2016. 76(2): p. 215-225.
  2. Iguchi, K., et al., Isolation and characterization of LNCaP sublines differing in hormone sensitivity. Journal of andrology, 2007. 28(5): p. 670-678.
  3. Wei, C., et al., CRISPR/Cas9 targeting of the androgen receptor suppresses the growth of LNCaP human prostate cancer cells. Molecular medicine reports, 2018. 17(2): p. 2901-2906.
  4. Gomes, I.M., et al., Knockdown of STEAP1 inhibits cell growth and induces apoptosis in LNCaP prostate cancer cells counteracting the effect of androgens. Medical Oncology, 2018. 35: p. 1-10.
  5. Tousi, M.S., et al., Evaluation of apoptotic effects of mPEG-b-PLGA coated iron oxide nanoparticles as a eupatorin carrier on DU-145 and LNCaP human prostate cancer cell lines. Journal of Pharmaceutical Analysis, 2021. 11(1): p. 108-121.