Şimay Demir Yaprak Dilber, Mohammed Ahmed Islam, Özdemir Aysun, Ark Mustafa
Department of Pharmacology, Faculty of Pharmacy, Gazi University, Ankara, Türkiye.
Med Oncol. 2025 Sep 17;42(11):475. doi: 10.1007/s12032-025-03030-7.
Caveolae and Rho kinase signaling individually are both remodeled during cellular senescence and influence cell morphology and SASP secretion. However, how this interaction modulates senescent cancer-cell morphology and SASP remains unresolved. We evaluated the possible connection between caveolae and ROCK on the development of senescent cell morphology and the secretion of the SASP.
Senescence was induced in HeLa and A549 cells with ouabain. Caveolin depletion was achieved with methyl-beta-cyclodextrin (MβCD) or Caveolin-1 siRNA applications. Cell morphology was assessed by cell area, volume, and thickness parameters, while SASP secretion was assessed by measuring total protein, IL-6, and VEGF-A in senescent cell secretomes. We also evaluated the effect of caveolae depletion on ROCK expression, activation, and localization.
While the caveolae disruption with the preincubation of MβCD did not alter the occurrence of ouabain-induced senescence, it significantly altered cellular morphological features, such as decreased cell area and volume. The amount of ROCK1 and ROCK2 in the membrane fraction of cells was decreased by MβCD preincubation. These findings indicate that there may be a relationship between ROCK and the morphological changes observed in senescent cells as a result of the disruption of the caveolar structure. However, the incubation of MβCD and caveolin-1 knocking down increased the secretory activity of senescent cells. Overall secretion increases with caveolar depletion, indicating that its secretion-enhancing effect is greater than any concomitant decrease that may occur with ROCK inhibition.
These data suggest that maintaining the integrity of caveolar structures plays a critical role in mitigating the detrimental effects of SASP released from chemotherapy-induced senescent cells.
小窝和Rho激酶信号传导在细胞衰老过程中各自都会发生重塑，并影响细胞形态和衰老相关分泌表型（SASP）的分泌。然而，这种相互作用如何调节衰老癌细胞的形态和SASP仍未得到解决。我们评估了小窝与Rho相关卷曲螺旋蛋白激酶（ROCK）之间在衰老细胞形态发展和SASP分泌方面的可能联系。
用哇巴因诱导HeLa和A549细胞衰老。通过应用甲基-β-环糊精（MβCD）或小窝蛋白-1（Caveolin-1）小干扰RNA（siRNA）来实现小窝蛋白的缺失。通过细胞面积、体积和厚度参数评估细胞形态，同时通过测量衰老细胞分泌组中的总蛋白、白细胞介素-6（IL-6）和血管内皮生长因子-A（VEGF-A）来评估SASP分泌。我们还评估了小窝缺失对ROCK表达、激活和定位的影响。
虽然用MβCD预孵育破坏小窝并没有改变哇巴因诱导的衰老的发生，但它显著改变了细胞形态特征，如细胞面积和体积减小。MβCD预孵育使细胞膜部分中ROCK1和ROCK2的量减少。这些发现表明，ROCK与由于小窝结构破坏而在衰老细胞中观察到的形态变化之间可能存在关系。然而，MβCD孵育和小窝蛋白-1敲低增加了衰老细胞的分泌活性。随着小窝缺失，总体分泌增加，表明其分泌增强作用大于ROCK抑制可能伴随的任何减少。
这些数据表明，维持小窝结构的完整性在减轻化疗诱导的衰老细胞释放的SASP的有害影响方面起着关键作用。
