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SiHa Cells - Shedding Light on Cervical Cancer Research

SiHa cells are cervical cancer cells widely used in biomedical research. They are great transfection hosts and, thus are suitable for gene expression studies. Due to their human origin and cervical cancer relevance, these cells are mainly employed in cancer biology, virology, and drug development studies.

This article aims to elucidate the basic information about cervical cancer SiHa cells that will help your working with it. You will learn:

General characteristics and origin of SiHa cells

Understanding a cell line's origin and general attributes is crucial, as it influences its use within your research. In this article section, you will learn about SiHa cell line origin, distinctive characteristics, and much more: What is the SiHa cancer cell line? What are SiHa cells HPV? What is the SiHa cell line origin? What is the morphology of SiHa?

  • SiHa, the cervical carcinoma cell line was derived from fragments of a primary uterine biopsy of an Asian female (55-year-old) with squamous cell cancer [1].
  • SiHa cell line displays a hypertriploid karyotype. Most of the cells have an average chromosome number between 69 and 72. However, around 24% of cells possess a modal chromosome number of 71.
  • SiHa cells are human papilloma virus 16 positive. They exhibit integration of around 1 to 2 copies of the HPV genome per cell [2].
  • These cervical cancer cells exhibit an epithelial morphology.

Division of HeLa cervical cancer cells under the microscope.

SiHa cell line: Culturing information

Maintaining a cell line culture is not easy until you know all the key points for culturing it. You should know: What is the doubling time of SiHa? What is SiHa cell media? How do you culture the SiHa cell line?

Key Points for Culturing SiHa Cells

Doubling Time:

The doubling time for SiHa cells is approximately 17 hours.

Adherent or in Suspension:

SiHa is an adherent cell line.

Sub-cultivation Ratio:

The subcultivation ratio for SiHa cells is 1:2 to 1:4. For passaging, and adherent cells are rinsed with 1x PBS. Accutase solution is added, and cells are incubated for 8 to 10 minutes at ambient temperature. Next, culture media is added, and cells are centrifuged. The cell pellet is resuspended, and cells are poured into the new flask for culturing.

Growth Medium:

Eagle's minimum essential medium (EMEM) containing 10% FBS, 2 mM L-Glutamine, 2.2 g/L NaHCO3, and Earle's Balanced Salt Solution (EBSS) is used for the ideal growth of SiHa cells. Media is replaced 2 to 3 times per week.

Growth Conditions:

SiHa cells are maintained in a 37°C humidified incubator with a 5% CO2 supply.

Storage:

The frozen SiHa cervical cancer cell line vials are stored at below -150°C temperature in an electric freezer or the vapour phase of liquid nitrogen for a longer term.

Freezing Process and Medium:

CM-1 or CM-ACF freezing medium is used for SiHa cell freezing. Cells are frozen using a slow freezing method that allows only a 1 °C decrease per minute to protect the viability of cells.

Thawing Process:

Frozen cells are kept for 40 to 60 seconds in a 37°C water bath until a small ice clump is left. Afterwards, fresh growth media is added, and cells are centrifuged to remove freezing media components. The cell pellet is resuspended, and cells are dispensed into a culture flask for growth.

Biosafety Level:

Biosafety level 1 laboratory is required to handle and maintain SiHa cell cultures.

An adherent layer of SiHa cells at 10× and 20× magnification.

Advantages & Disadvantages of SiHa cell line

Like other human carcinoma cell lines, SiHa also possesses some distinct characteristics associated with certain advantages and drawbacks. Herein, we have discussed a few significant ones.

Advantages of the SiHa Cervical Cancer Cell Line

  • Cervical Cancer Model
    • Derived from cervical squamous cell carcinoma.
    • Used to study the mechanisms, growth, and development of cervical cancer.
    • Expresses genes p53+ and pRB+ which are related to DNA repair, cell cycle regulation, and tumor suppression.
  • Tumorigenic Potential
    • SiHa cells are tumorigenic and can induce poorly differentiated grade III epidermoid tumors in nude mice.
    • Used for in vivo cancer research and testing anti-cancer treatments.

Disadvantages of SiHa Cells

  • Proliferation Rate
    • SiHa cells proliferate rapidly, leading to overgrowth.
    • Requires frequent passaging, increasing the risk of genetic instability, which may impact cell behavior over time.

Research applications of SiHa cells

The SiHa cell line is extensively used in cervical cancer research. A few specified applications of this cell line are discussed here.

  • Human papillomavirus (HPV) related studies: SiHa cells are HPV 16 positive thus, they are valuable entities for studying human papillomavirus infection, its molecular mechanisms, and its role in the development and progression of cervical cancer. Researchers also employ these cells to investigate viral replication, integration, and its impact on host cell processes. A study conducted in 2020 used the SiHa cell line to examine the role of HPV E6/E7 oncoproteins in cervical cancer development by targeting it with CRISPR technology. The findings revealed that the knockdown of E6/E7 genes caused SiHa cell growth inhibition and apoptosis. Therefore, these viral genes may serve as crucial drug targets for developing therapies against HPV-associated cervical cancer [3].
  • Cancer biology: SiHa cells serve as an invaluable model for studying cervical cancer biology, including cancer development, progression, metastasis, and invasion. Researchers utilize these cells to explore the genetic mutations and underlying molecular pathways contributing to cervical cancer development and growth, such as, a study utilized SiHa cells and found that upregulated SEC61G (SEC61 translocon subunit gamma) encourages cervical cancer cell proliferation through activation of MAPK cascade [4].
  • Drug screening and testing: SiHa is a widely used cervical cancer line to assess the efficacy of potential anti-cancer drugs specific to HPV-associated cervical cancer. Researchers explore the antiproliferative, apoptotic, and anti-metastatic potential of drugs using these cells. Such as a study carried out in 2019 that investigated the cytotoxic potential of Vatica diospyroides Symington Type SS fruit extracts on SiHa cervical cancer cell line [5].

SiHa cells for your research!

Research Publications Featuring SiHa Cells

Some significant and frequently cited research publications are featuring the SiHa cervical squamous carcinoma cell line.

Anti-tumor effect of aloe-emodin on cervical cancer cells was associated with human papillomavirus E6/E7 and glucose metabolism

This publication in OncoTargets and Therapy (2019) proposed that a natural compound aloe-emodin exerts HPV E6/E7 and glucose metabolism-associated apoptotic effects in SiHa cervical cancer cells.

Evaluation of anti-cancer potential of Excoecaria agallocha (L.) leaf extract on human cervical cancer (SiHa) cell line and assessing the underlying mechanism of action

This article was published in the Future Journal of Pharmaceutical Sciences in 2022. This study investigated the anti-tumor activities of Excoecaria agallocha (L.) plant leaf extract in the SiHa cervical cancer cell line.

Antiproliferative effect of 1, 10-Phenanthroline coupled to sulfated ZnO nanoparticles in SiHa cervix cancer cell line

This study published in the Journal of Sol-Gel Science and Technology (2022) proposed that the 1, 10-Phenanthroline molecule upon coupling with zinc sulfate nanoparticles, exerts significant antiproliferative effects in SiHa cells.

Astragaloside IV inhibits the invasion and metastasis of SiHa cervical cancer cells via the TGF‑β1‑mediated PI3K and MAPK pathways

This research was published in the Oncology Reports in 2019. The study revealed that Astragaloside IV, a natural product, suppresses the migration and invasion of SiHa cells by modulating TGF‑β1‑mediated PI3K and MAPK signalling pathways.

IFI16 promotes cervical cancer progression by upregulating PD-L1 in immunomicroenvironment through STING-TBK1-NF-kB pathway

This study in Biomedicine & Pharmacotherapy (2020) revealed that IFI16 (interferon-gamma inducible protein 16) regulates the PD-L1 gene via activating the STING-TBK1-NF-kB pathway to encourage SiHa cervical cancer cell growth.

Resources for SiHa Cell line: Protocols, Videos, and More

Here are some online resources featuring the SiHa cells.

  • SiHa cell transfection: This research article contains a protocol for maintaining SiHa cervical cancer cells and for SiHa cell transfection.

The following link contains cell culture information for SiHa cells. 

  • SiHa cell line: This website contains much valuable data about the SiHa cell line. This includes information about growth media, culturing conditions, protocols for subculturing SiHa cells and handling its proliferative and cryopreserved cultures.

FAQs for SiHa Cervical Carcinoma Cells

SiHa cells are a human cancer cell line derived from cervical squamous cell carcinoma. They are extensively used in research for cervical carcinomas, particularly in studies related to human papillomavirus type 16 (HPV-16), known to play a crucial role in the pathogenesis of cervical cancer.

In SiHa cells, HPV-16 DNA is typically integrated into the host cell's chromosomes. Chromosomal integration sites of HPV can be critical in understanding the progression and treatment resistance of cervical carcinoma.

Yes, SiHa cells are a critical component of the cervical cancer cell panel utilized for the biological characterization of tumor cells, examining the effects of chemotherapeutics, and the development of new drugs for scientific research.

Absolutely. SiHa cells, which contain integrated HPV DNA, are invaluable for in situ hybridization and other molecular biology techniques to study the role of HPV in the development of cervical neoplasms and the potential enhancement of curcumin effects on cancer treatment.

SiHa cells are tumorigenic in immunodeficient mice, making them a suitable model for studying the pathogenicity of cervical carcinoma and the in vivo effects of potential therapeutics.

References for SiHa cells can be found in scientific literature across journals such as Cancer Research (Cancer Res), Biochemistry and Cell Biology, Journal of Cellular Physiology, and more. They provide detailed information on the use of SiHa cells in the context of cancer genome studies, phenotyping, and the examination of gene functions.

The genetic profile of SiHa cells, including the expression of several genes and the acquisition of CDV, contributes to the comprehensive understanding of cervical carcinoma's etiology and the development of targeted therapies.

SiHa cells are maintained in primary cell culture, adhering to specific growth conditions and medium requirements to preserve their characteristics and functionality for research purposes.

References

  1. Melzer, C., J. von der Ohe, and R. Hass, Concise review: crosstalk of mesenchymal stroma/stem-like cells with cancer cells provides therapeutic potential. Stem cells, 2018. 36(7): p. 951-968.
  2. Ostrowska, K.M., et al., Investigation of the influence of high-risk human papillomavirus on the biochemical composition of cervical cancer cells using vibrational spectroscopy. Analyst, 2010. 135(12): p. 3087-3093.
  3. Chen, Y., et al., In vitro and in vivo growth inhibition of human cervical cancer cells via human papillomavirus E6/E7 mRNAs’ cleavage by CRISPR/Cas13a system. Antiviral Research, 2020. 178: p. 104794.
  4. Fan, Y., et al., SEC61G promotes cervical cancer proliferation by activating MAPK signaling pathway. Disease Markers, 2022. 2022.
  5. Chothiphirat, A., et al., Anticancer potential of fruit extracts from Vatica diospyroides symington type SS and their effect on program cell death of cervical cancer cell lines. The Scientific World Journal, 2019. 2019.

 

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