Aksoy Asude, Göktürk Selcen, Etem Önalan Ebru, Tektemur Ahmet, Artaş Gökhan, Varoğlu Asuman, Koç Mustafa
Department of Medical Oncology, Medical Faculty, Firat University, Elazig, Turkey.
Department of Internal Medicine, Medical Faculty, Firat University, Elazig, Turkey.
Turk J Biol. 2022 May 31;46(3):239-250. doi: 10.55730/1300-0152.2612. eCollection 2022.
Stathmin1 (STMN1) has been proposed as a possible prognostic marker and a potential therapeutic target for some cancers. We aimed to analyze the changes in autophagy, invasion, apoptosis-related genes in prostate cancer (PCa) cell line (PC-3), after small interfering RNA (siRNA)-mediated STMN1 silencing, and also the relationships of STMN1 expression, clinicopathological parameters, and survival (OS) in PCa cases. The STMN1 expressions were analyzed, immunohistochemically, in formalin-fixed paraffin-embedded 75 PCa and 15 benign prostatic hypertrophy (BPH) tissues. The correlation between the levels of expression STMN1, clinicopathological features, and OS was determined in PCa cases. The siRNA-mediated STMN1 incubated PC-3 cells were transfected and compared to negative control siRNAs. We determined mRNA levels in autophagy, invasion, and apoptosis genes with the combination of reverse transcription-polymerase chain reaction (RT-PCR) and western blotting in PC3 cell lines after STMN1 silencing. It was determined that STMN1 was overexpressed significantly in PCa cases, immunohistochemically. The overexpression of STMN1 was significantly correlated with the high-grade Gleason score, and it was associated with a worse prognosis of PCa cases according to the Kaplan-Meier survival analysis (p < 0.05). Significant silencing in STMN1 was determined (87.5%) after siRNA applications. Especially, invasion genes such as claudin 7, fibroblast growth factor 8, hypoxia-inducible factor 1 subunit alpha, hepatocyte growth factor, matrix metallopeptidase 2, 7 genes, markedly, decreased by siRNA-mediated STMN1silencing. STMN1 silencing was determined to significantly increase caspase 3 protein expression by using western blot analysis (p < 0.001). Although STMN1 silencing did not have a significant effect on the induction of apoptosis and autophagy-related genes in PCa cells, it was shown to affect apoptotic mechanisms through the caps3 protein. siRNA-mediated STMN1 silencing decreases proliferation in the PCa cell line. It is thought that STMN1 can serve as a potential therapeutic target in the advanced stage-PCa, especially.
Stathmin1(STMN1)已被提议作为某些癌症可能的预后标志物和潜在治疗靶点。我们旨在分析小干扰RNA(siRNA)介导的STMN1沉默后前列腺癌细胞系(PC-3)中自噬、侵袭、凋亡相关基因的变化,以及前列腺癌(PCa)病例中STMN1表达、临床病理参数和生存率(OS)之间的关系。采用免疫组织化学方法分析了75例PCa和15例良性前列腺增生(BPH)组织标本(经福尔马林固定、石蜡包埋)中STMN1的表达情况。确定了PCa病例中STMN1表达水平、临床病理特征和OS之间的相关性。将经siRNA介导的STMN1孵育的PC-3细胞进行转染,并与阴性对照siRNA进行比较。在STMN1沉默后的PC3细胞系中,我们结合逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法测定了自噬、侵袭和凋亡基因的mRNA水平。免疫组织化学检测发现,PCa病例中STMN1明显过表达。根据Kaplan-Meier生存分析,STMN1的过表达与高分级Gleason评分显著相关,且与PCa病例的预后较差有关(p<0.05)。应用siRNA后,STMN1的表达被显著沉默(87.5%)。特别是,siRNA介导的STMN1沉默显著降低了claudin 7、成纤维细胞生长因子8、缺氧诱导因子1α亚基、肝细胞生长因子、基质金属蛋白酶2等侵袭相关基因的表达水平。蛋白质免疫印迹分析显示,STMN1沉默可显著增加caspase 3蛋白的表达(p<0.001)。虽然STMN1沉默对PCa细胞中凋亡和自噬相关基因的诱导没有显著影响,但它可通过casp3蛋白影响凋亡机制。siRNA介导的STMN1沉默可降低PCa细胞系中的增殖。尤其值得一提的是,STMN1有望成为晚期PCa的潜在治疗靶点。