背景:本研究调查了他扎罗汀诱导基因1(TIG1)抑制黑色素瘤细胞生长的机制。主要重点是分析黑色素瘤细胞中TIG1调控的下游基因及其对细胞生长的影响。
方法:使用水溶性四唑1(WST-1)线粒体染色和乳酸脱氢酶释放测定评估TIG1表达对细胞活力和死亡的影响。RNA测序和蛋白质印迹分析用于研究黑色素瘤细胞中TIG1调控的基因。此外,在黑色素瘤组织阵列中分析了TIG1表达与其下游基因之间的相关性.
结果:黑色素瘤细胞中TIG1的表达与细胞活力降低和细胞死亡增加相关。RNA测序(RNA-seq),定量反转录PCR(反向RT-QPCR),和免疫印迹显示TIG1表达诱导内质网(ER)应激反应相关基因的表达,例如同型半胱氨酸反应性内质网驻留泛素样结构域成员1(HERPUD1),结合免疫球蛋白(BIP),和DNA损伤诱导型转录物3(DDIT3)。此外,黑色素瘤组织阵列分析显示TIG1表达与HERPUD1、BIP、DDIT3此外,黑色素瘤细胞内质网应激反应的减弱削弱了TIG1对细胞生长的影响。
结论:TIG1表达有效阻碍黑色素瘤细胞的生长。TIG1诱导ER应激反应相关基因的上调,导致caspase-3活性增加和随后的细胞死亡。这些发现表明,视黄酸预防黑色素瘤形成的能力可能与TIG1的抗癌作用有关。
BACKGROUND: This study investigated the mechanism by which tazarotene-induced gene 1 (TIG1) inhibits
melanoma cell growth. The main focus was to analyze downstream genes regulated by TIG1 in
melanoma cells and its impact on cell growth.
METHODS: The effects of TIG1 expression on cell viability and death were assessed using water-soluble tetrazolium 1 (WST-1) mitochondrial staining and lactate dehydrogenase release assays. RNA sequencing and Western blot analysis were employed to investigate the genes regulated by TIG1 in melanoma cells. Additionally, the correlation between TIG1 expression and its downstream genes was analyzed in a melanoma tissue array.
RESULTS: TIG1 expression in
melanoma cells was associated with decreased cell viability and increased cell death. RNA-sequencing (RNA-seq), quantitative reverse transcription PCR (reverse RT-QPCR), and immunoblots revealed that TIG1 expression induced the expression of Endoplasmic Reticulum (ER) stress response-related genes such as Homocysteine-responsive endoplasmic reticulum-resident ubiquitin-like domain member 1 (HERPUD1), Binding immunoglobulin protein (BIP), and DNA damage-inducible transcript 3 (DDIT3). Furthermore, analysis of the melanoma tissue array revealed a positive correlation between TIG1 expression and the expression of HERPUD1, BIP, and DDIT3. Additionally, attenuation of the ER stress response in
melanoma cells weakened the impact of TIG1 on cell growth.
CONCLUSIONS: TIG1 expression effectively hinders the growth of
melanoma cells. TIG1 induces the upregulation of ER stress response-related genes, leading to an increase in caspase-3 activity and subsequent cell death. These findings suggest that the ability of retinoic acid to prevent melanoma formation may be associated with the anticancer effect of TIG1.