Posters

Presenting Author Academic/Professional Position

Veronica O'Brien, Medical Student

Academic Level (Author 1)

Medical Student

Academic Level (Author 2)

Faculty

Discipline/Specialty (Author 2)

Cancer and Immunology

Academic Level (Author 3)

Faculty

Discipline/Specialty (Author 3)

Cancer and Immunology

Academic Level (Author 4)

Faculty

Discipline/Specialty (Author 4)

Cancer and Immunology

Academic Level (Author 5)

Faculty

Discipline/Specialty (Author 5)

Internal Medicine

Discipline Track

Community/Public Health

Abstract Type

Research/Clinical

Abstract

Background: Cervical cancer is the 4th leading cause of cancer-related death worldwide. Cervical cancer is primarily caused by high-risk strains of the HPV virus. HPV upregulates oncoproteins E6 and E7, disrupting p53 and pRb functions. Several databases, including the Human Genome Atlas, Tumor Immune Estimation Resource, UALCAN, and COSMIC show increased SLC1A5 expression in cervical cancer. SLC1A5, also known as the alanine-serine-cysteine transporter, is a major transporter of glutamine and is found on the cell membrane. Our goal is to investigate the role of SLC1A5 in cervical cancer.

Methods: SLC1A5 mRNA expression in cervical cancer was examined using the Cancer Genome Atlas (TCGA), the Genotype Tissue Expression Atlas (GTEx), the Tumor Immune Estimation Resource (TIMER), and the UALCAN databases to examine the relationship between SLC1A5 expression and the clinicopathological characteristics of cervical cancer. We investigated the expression of SLC1A5 in various cervical cell lines through RT-PCR, western blot, and confocal imaging. HeLa cells contain two transcripts from the HPV18 virus, SiHa has one to two transcripts of the HPV16 virus, CaSki has 600 transcripts from the HPV16 virus, C-33a is a cervical cancer cell line that is not infected with HPV. We also investigated the expression of SLC1A5 in noncancerous, uninfected cervical cells using the CEC cell line.

Results: The SLC1A5 mRNA expression was upregulated in cervical cancer samples as compared to normal cervical tissues. For the cancer stages, ages, races, patients’ weight, histopathological subtype, and nodal metastasis status there existed significant difference in the expression of SLC1A5 among different groups by mining of the UALCAN database. The mutation landscape analysis confirms that SLC1A5 genetic alterations reached 3%, of which missense substitutions accounted for the highest proportion of 48.77%. The findings of the TIMER analysis indicated a correlation between immune cell infiltration and SLC1A5 copy number alterations (CNA). The expression level of SLC1A5 was positively correlated with the infiltration level of CD4+ T cells, neutrophils, and dendritic cells but negatively correlated with CD8+ T cells, B cells and macrophage infiltration. Additionally, SLC1A5 expression was also found to be associated with certain immunosuppressive cells, chemokines, and receptors. RNA expression of SLC1A5 in HeLa, SiHa, CaSki, C-33a and CEC, each with an n=3, was quantified via RTPCR. Tukey HSD test showed that CaSki had significantly higher levels of SLC1A5 than all the other cell lines (p-value < .01). While CEC had the lower expression of SLC1A5 compared to CaSki (p-value < .01) and SiHa (p-value < .01). Western blot analysis shows CaSki has the highest protein expression of SLC1A5 while CEC has significantly less expression. Of the cancer cell lines, C33-a has the lowest expression. Western blot densitometry quantification reflects the results from RT-PCR. Confocal microscopy also reflected the difference of SLC1A5 expression in CEC compared to other cell lines.

Conclusions: Our observation demonstrates that SLC1A5 was highly expressed in cervical cancer tissues and is linked to prognosis of cervical cancer patients and tumor immune responses. Experimental analysis shows that SLC1A5 expression is higher in cervical cell lines with increased levels of HPV transcripts

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SLC1A5 as a Prognostic Biomarker: Insights from Transcriptomic and Clinical Data in Cervical Cancer

Background: Cervical cancer is the 4th leading cause of cancer-related death worldwide. Cervical cancer is primarily caused by high-risk strains of the HPV virus. HPV upregulates oncoproteins E6 and E7, disrupting p53 and pRb functions. Several databases, including the Human Genome Atlas, Tumor Immune Estimation Resource, UALCAN, and COSMIC show increased SLC1A5 expression in cervical cancer. SLC1A5, also known as the alanine-serine-cysteine transporter, is a major transporter of glutamine and is found on the cell membrane. Our goal is to investigate the role of SLC1A5 in cervical cancer.

Methods: SLC1A5 mRNA expression in cervical cancer was examined using the Cancer Genome Atlas (TCGA), the Genotype Tissue Expression Atlas (GTEx), the Tumor Immune Estimation Resource (TIMER), and the UALCAN databases to examine the relationship between SLC1A5 expression and the clinicopathological characteristics of cervical cancer. We investigated the expression of SLC1A5 in various cervical cell lines through RT-PCR, western blot, and confocal imaging. HeLa cells contain two transcripts from the HPV18 virus, SiHa has one to two transcripts of the HPV16 virus, CaSki has 600 transcripts from the HPV16 virus, C-33a is a cervical cancer cell line that is not infected with HPV. We also investigated the expression of SLC1A5 in noncancerous, uninfected cervical cells using the CEC cell line.

Results: The SLC1A5 mRNA expression was upregulated in cervical cancer samples as compared to normal cervical tissues. For the cancer stages, ages, races, patients’ weight, histopathological subtype, and nodal metastasis status there existed significant difference in the expression of SLC1A5 among different groups by mining of the UALCAN database. The mutation landscape analysis confirms that SLC1A5 genetic alterations reached 3%, of which missense substitutions accounted for the highest proportion of 48.77%. The findings of the TIMER analysis indicated a correlation between immune cell infiltration and SLC1A5 copy number alterations (CNA). The expression level of SLC1A5 was positively correlated with the infiltration level of CD4+ T cells, neutrophils, and dendritic cells but negatively correlated with CD8+ T cells, B cells and macrophage infiltration. Additionally, SLC1A5 expression was also found to be associated with certain immunosuppressive cells, chemokines, and receptors. RNA expression of SLC1A5 in HeLa, SiHa, CaSki, C-33a and CEC, each with an n=3, was quantified via RTPCR. Tukey HSD test showed that CaSki had significantly higher levels of SLC1A5 than all the other cell lines (p-value < .01). While CEC had the lower expression of SLC1A5 compared to CaSki (p-value < .01) and SiHa (p-value < .01). Western blot analysis shows CaSki has the highest protein expression of SLC1A5 while CEC has significantly less expression. Of the cancer cell lines, C33-a has the lowest expression. Western blot densitometry quantification reflects the results from RT-PCR. Confocal microscopy also reflected the difference of SLC1A5 expression in CEC compared to other cell lines.

Conclusions: Our observation demonstrates that SLC1A5 was highly expressed in cervical cancer tissues and is linked to prognosis of cervical cancer patients and tumor immune responses. Experimental analysis shows that SLC1A5 expression is higher in cervical cell lines with increased levels of HPV transcripts

 

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