Relative to its incidence, oral cancer has serious negative social effects. The exact causes of oral cancer have not been clarified, but many studies have implicated smoking and drinking. However, the fundamental mechanism of oral cancer causation has yet to be elucidated. Lysophosphatidic acid (LPA...
Relative to its incidence, oral cancer has serious negative social effects. The exact causes of oral cancer have not been clarified, but many studies have implicated smoking and drinking. However, the fundamental mechanism of oral cancer causation has yet to be elucidated. Lysophosphatidic acid (LPA) augments epithelial mesenchymal transition (EMT) and development of various cancer cells. However, a detailed mechanistic explanation for LPA-induced EMT and the effects of EMT-promoting conditions on oral squamous cell carcinoma development remain elusive. In the present study, a quantitative reverse transcription polymerase chain reaction was used to analyze TWIST1, Slug, E-cadherin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcript expression. Immunoblotting was used to analyze TWIST1, Slug, E-cadherin, and GAPDH protein expression. siRNAs were used to silence TWIST1 and Slug transcript expression. A matrigel-coated in vitro invasion insert was used to analyze oral cancer cell invasion. The results of the present study show that the expression levels of TWIST1 and Slug, which are EMT factors, were increased by LPA treatment in YD-10B oral squamous cell carcinoma. Conversely, E-cadherin expression was significantly reduced. In addition, transfection of the cells with TWIST1 and Slug siRNA strongly inhibited LPA-induced oral cancer cell invasion. The present study shows that TWIST1 and Slug mediate LPA-induced oral cancer cell EMT and invasiveness. The present study confirmed the mechanism by which LPA promotes oral cancer cell development, with TWIST1 and Slug providing novel biomarkers and promising therapeutic targets for oral cancer cell development.
Relative to its incidence, oral cancer has serious negative social effects. The exact causes of oral cancer have not been clarified, but many studies have implicated smoking and drinking. However, the fundamental mechanism of oral cancer causation has yet to be elucidated. Lysophosphatidic acid (LPA) augments epithelial mesenchymal transition (EMT) and development of various cancer cells. However, a detailed mechanistic explanation for LPA-induced EMT and the effects of EMT-promoting conditions on oral squamous cell carcinoma development remain elusive. In the present study, a quantitative reverse transcription polymerase chain reaction was used to analyze TWIST1, Slug, E-cadherin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcript expression. Immunoblotting was used to analyze TWIST1, Slug, E-cadherin, and GAPDH protein expression. siRNAs were used to silence TWIST1 and Slug transcript expression. A matrigel-coated in vitro invasion insert was used to analyze oral cancer cell invasion. The results of the present study show that the expression levels of TWIST1 and Slug, which are EMT factors, were increased by LPA treatment in YD-10B oral squamous cell carcinoma. Conversely, E-cadherin expression was significantly reduced. In addition, transfection of the cells with TWIST1 and Slug siRNA strongly inhibited LPA-induced oral cancer cell invasion. The present study shows that TWIST1 and Slug mediate LPA-induced oral cancer cell EMT and invasiveness. The present study confirmed the mechanism by which LPA promotes oral cancer cell development, with TWIST1 and Slug providing novel biomarkers and promising therapeutic targets for oral cancer cell development.
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제안 방법
PCR amplification was conducted using the following specific primer sets: TWIST1: forward 5’-GTC CGC AGT CTT ACG AGG AG-3’/reverse 5’-CCA GCT TGA GGG TCT GAA TC-3’; Slug: forward 5’-TCT GCA GAC CCA TTC TGA TG-3’/5’-AGC AGC CAG ATT CCT CAT GT-3’; E-cadherin: forward 5’-ACA GCC CCG CCT TAT GAT T-3’/5’-TCG GAA CCG CTT CCT TCA-3’; GAPDH: forward 5’-ACA GTC AGC CGC ATC TTC TT-3’/reverse, 5’-ACG ACC AAA TCC GTT GAC TC-3’.
In order to investigate the effects of LPA on expression of EMT factors, we treated YD-10B cells with LPA at different times, and measured the mRNA (Fig. 1A) and protein (Fig. 1B) expression of TWIST1, Slug, and E-cadherin, which are known to be important factors in EMT. We found that mRNA expression was the highest for TWIST1 at 12 hours and for Slug at 2 hours (Fig.
이론/모형
GAPDH was used as a control for calculating dCt. The RT-PCR data were analyzed using the 2-(ddCt) method.
성능/효과
In the present study, we verified LPA-induced EMT and invasion effects in oral cancer cells. Our results demonstrate not only that LPA causes an increase in expression of 2 important EMT factors, TWIST1 and Slug in oral cancer, but also that silencing TWIST1 and Slug caused a significant decrease in LPA-induced invasion by oral cancer cells. This means that TWIST1 and Slug are important biomarkers for LPA-induced oral cancer cell invasion.
후속연구
Our study is important because it investigates the effects of LPA on oral cancer cell invasion. However, because the experiments were conducted at the cell level, additional animal experiments and clinical studies will be required. If a drug were developed that could suppress mechanisms of LPA-induced oral cancer cell metastasis, we expect it would show a revolutionary inhibitory effect on oral cancer.
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