Page 11 - EJMO-9-1

Eurasian Journal of Medicine and
Oncology
Single-cell RNA-seq in malignant skin tumors

lineage features in the human epidermis and providing platforms for large-scale, parallel sequencing.
a multidimensional characterization of inflammatory Common platforms include Illumina and MGI,
diseases. Moreover, the findings also demonstrated the producing transcriptome data for each individual cell.
potential to classify skin rashes of previously undetermined This article provides a concise overview of the single-
origin, underscoring the significant role of single-cell cell sequencing analysis process, emphasizing the latest
technology in clinical applications. research advancements in single-cell transcriptome
In 2009, Tang et al. performed the first single-cell sequencing aimed at characterizing heterogeneity,
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sequencing of the entire transcriptome. Since then, reshaping the microenvironment, and understanding
various improvements and new technologies, such as resistance mechanisms in three major skin cancers. Our
SMART-seq, Drop-seq, and 10× Genomics, have driven review serves as a stepping stone toward developing
the widespread adoption of single-cell sequencing. innovative therapeutic strategies and personalized
Today, single-cell sequencing technology has expanded treatment approaches.
beyond RNA-seq to encompass DNA sequencing,
protein sequencing, epigenomic sequencing, and spatial 3. The research progress of single-cell RNA-
transcriptomics, among other multi-omics techniques. seq in BCC
These multidimensional data provide researchers with a 3.1. The main clinical features of BCC
more comprehensive understanding of cellular biological
characteristics. BCC ranks among the most prevalent malignant skin
tumors, originating from basal cells. It primarily affects
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The procedure for single-cell transcriptome sequencing elderly individuals and commonly manifests on the
mainly involves three key steps: (i) single-cell isolation, head and face. Despite its typically slow growth and low
(ii) library preparation, and (iii) sequencing (Figure 2). malignant potential, the incidence of BCC continues to
(i) Single-cell isolation: Common methods for isolating rise. Initially asymptomatic, it can progressively lead to
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single cells include fluorescence-activated cell sorting, local tissue destruction and recurrent ulceration. Early
manual sorting under fluorescence microscopy, intervention significantly enhances prognosis; however,
microfluidic technologies, centrifugal separation, untreated cases may result in disfiguring injuries and
flow cytometry sorting, magnetic bead separation, potential infiltration of surrounding tissues, often with
microfluidic systems, and gel bead methods. Among high recurrence rates. Prolonged exposure to sunlight
these, microfluidic technologies, which use tiny fluid and UV radiation, which can damage DNA structures, is
channels and control systems to isolate individual cells a probable etiological factor in the development of BCC.
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and distribute them into small units, are widely utilized Clinically and histopathologically, BCC is categorized into
for their efficiency and cost-effectiveness in managing several types, including nodular-ulcerative, superficial,
large volumes of cells. The choice of single-cell isolation cystic, fibroepithelial, and infiltrative BCC, with nodular-
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method should be guided by specific experimental ulcerative being the most prevalent. Diagnostic methods
requirements, such as sample type, cell quantity, include dermoscopy, reflectance confocal microscopy, and
experimental goals, and the availability of equipment optical coherence tomography. The primary treatment
and technology. Combining various methods can for BCC involves surgical excision, complemented by
enhance the accuracy and efficiency of cell sorting. alternative options such as laser therapy, radiotherapy,
(ii) Library preparation: Once isolated, single cells immunotherapy, or targeted therapy.
undergo lysis to release RNA. RNA is transcribed into
complementary DNA for PCR amplification through 3.2. The complex regulatory mechanisms within the
reverse transcription. Sample barcodes and sequences tumor microenvironment (TME) of BCC
required for the Illumina sequencing platform are Recent advancements in single-cell transcriptome
added during amplification. Finally, the amplified sequencing technology have significantly contributed to
library undergoes fragment size selection and quality BCC research, uncovering the complex interplay between
control. Commercialized single-cell transcriptome immune cells, stromal cells, tumor cells, and the TME
sequencing technologies and platforms, such as during BCC initiation, progression, and treatment response.
10x Genomics Chromium System, BD Rhapsody, A study integrating in vivo optical coherence tomography,
Singleron Matrix, and Seekone DD, facilitate efficient scRNA-seq, spatial global transcriptional profiling, and
processing and library preparation. in situ sequencing has created comprehensive spatial maps
(iii) High-throughput sequencing: The prepared libraries of adult human skin across different anatomical sites and
are loaded onto high-throughput sequencing BCC at single-cell resolution. This study provides a detailed

Volume 9 Issue 1 (2025) 3 doi: 10.36922/ejmo.5809