Heat shock protein 27 (Hsp27) is an important member of the chaperone protein family and its overexpression promotes cancer cell survival. Here, we investigated the apoptosis inducer role of the J2 compound (Hsp27 inhibitor) in human ovarian cancer cell lines (SKOV3 and OVCAR-3). Cell proliferation was measured by MTT assay. The parameters of J2-Hsp27 interaction were determined with molecular docking calculation. The inhibitory effect of the J2 compound on Hsp27 chaperone activity was investigated by luciferase activity assay. Finally, the apoptotic inducer role of the J2 compound on SKOV3 and OVCAR-3 cells was determined by RT-PCR and caspase-3 activity assay. J2 compound decreased SKOV3 and OVCAR-3 cell proliferation in a dose-dependent manner at 48 h with IC₅₀ values of 17.34 µM and 12.63 µM, respectively. J2 inhibited the refolding process of denatured luciferase as an Hsp27 inhibitor. Molecular docking calculation was carried out to determine the interaction between Hsp27 and J2. The results indicated that J2 selectively binds to the phosphorylation site of the Hsp27 and inhibits the phosphorylation process of Hsp27. To determine the apoptotic potential of the J2 compound against ovarian cancer cells, the mRNA expression levels of apoptotic and antiapoptotic markers (Bax, Bcl-2, Bcl-xL, Cyt-c, p53, Apaf-1, Cas-3, Cas-8, Cas-9, TNF-α, DAXX, and Ask-1) were measured using RT-PCR. While J2 increased the expressions of apoptotic genes, it decreased the expressions of anti-apoptotic genes. Further, the J2 compound increased Cas-3 activity in SKOV3 and OVCAR-3 at 5.52 and 4.12 folds, respectively. These results confirm that J2 has great potential and significance in the stimulation of apoptosis in ovarian cancer cells as an Hsp27 inhibitor.

热休克蛋白 27 (Hsp27) 是伴侣蛋白家族的重要成员，其过度表达可促进癌细胞存活。在这里，我们研究了 J2 化合物（Hsp27 抑制剂）在人卵巢癌细胞系（SKOV3 和 OVCAR-3）中的细胞凋亡诱导作用。通过MTT测定来测量细胞增殖。通过分子对接计算确定J2-Hsp27相互作用的参数。通过荧光素酶活性测定研究J2化合物对Hsp27分子伴侣活性的抑制作用。最后，通过RT-PCR和caspase-3活性测定确定J2化合物对SKOV3和OVCAR-3细胞的凋亡诱导作用。J2化合物在48小时时以剂量依赖性方式降低SKOV3和OVCAR-3细胞增殖，IC ₅₀值分别为17.34 µM和12.63 µM。J2 作为 Hsp27 抑制剂抑制变性荧光素酶的重折叠过程。进行分子对接计算以确定Hsp27与J2之间的相互作用。结果表明J2选择性地结合Hsp27的磷酸化位点并抑制Hsp27的磷酸化过程。为了确定 J2 化合物对卵巢癌细胞的凋亡潜力，检测了凋亡和抗凋亡标志物（Bax、Bcl-2、Bcl-xL、Cyt-c、p53、Apaf-1、Cas-3、Cas-）的mRNA 表达水平。 8、Cas-9、TNF-α、DAXX和Ask-1）使用 RT-PCR 进行测量。J2增加了凋亡基因的表达，同时降低了抗凋亡基因的表达。此外，J2 化合物将 SKOV3 和 OVCAR-3 中的 Cas-3 活性分别提高了 5.52 倍和 4.12 倍。这些结果证实J2作为Hsp27抑制剂在刺激卵巢癌细胞凋亡方面具有巨大的潜力和意义。