Zhu Haiyan, Tang Yuxin, Zhang Xiangyang, Jiang Xianzhen, Wang Yong, Gan Yu, Yang Jianfu
Department of Anesthesiology, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China.
Med Oncol. 2015 Mar;32(3):84. doi: 10.1007/s12032-015-0541-y. Epub 2015 Feb 21.
Uroplakin 1A (UPK1A) is a specific marker of mammalian urothelium and one of major proteins contained in urothelial plaques. Many recent studies reported that UPK1A could be useful marker for diagnosis, detection and prognostic prediction of transitional cell carcinoma. However, relatively little is known about its exact roles in bladder transitional cell carcinoma (BTCC). We tried to explore the roles UPK1A plays in BTCC via the transfection of its antisense nucleotides (AS) into T24 cells to observe their changes of proliferation and apoptosis. After AS was successfully transfected into T24 cells, the percentages of proliferating T24 cells at 24 and 48 h after the treatment were 57.2 ± 6.8 and 44.7 ± 5.2%, significantly lower than that of control group, as shown by MTT (p < 0.05 and 0.01). At 24 h after transfection of AS, the percentage of apoptotic T24 cells was 26.87% measured by flow cytometry, significantly higher than that of control group (p < 0.01). Similarly, Hoechst 33258 staining showed that the percentage of apoptotic nuclei of T24 cells after 24 h treated by AS was 28.9%, significantly higher than that of control (p < 0.05). The most common and typical morphological changes of apoptosis, including shrink, pyknosis and karyorrhexis of T24 cells nuclei and DNA fragmentation were seen from Hoechst 33258 staining and DNA agarose gel electrophoresis. Taken together, inhibition of UPK1A can suppress proliferation and enhance apoptosis of BTCC T24 cells, suggesting it a potential target to treat this disease.
尿血小板溶素1A（UPK1A）是哺乳动物尿路上皮的特异性标志物，也是尿路上皮斑块中所含的主要蛋白质之一。最近的许多研究报道，UPK1A可能是诊断、检测和预测移行细胞癌的有用标志物。然而，关于其在膀胱移行细胞癌（BTCC）中的确切作用知之甚少。我们试图通过将其反义核苷酸（AS）转染到T24细胞中，观察其增殖和凋亡变化，来探索UPK1A在BTCC中的作用。将AS成功转染到T24细胞后，处理后24小时和48小时增殖的T24细胞百分比分别为57.2±6.8%和44.7±5.2%，显著低于对照组，MTT检测显示（p<0.05和0.01）。转染AS后24小时，流式细胞术检测凋亡的T24细胞百分比为26.87%，显著高于对照组（p<0.01）。同样，Hoechst 33258染色显示，AS处理24小时后T24细胞凋亡细胞核的百分比为28.9%，显著高于对照组（p<0.05）。从Hoechst 33258染色和DNA琼脂糖凝胶电泳中可以看到凋亡最常见和典型的形态学变化，包括T24细胞核的皱缩、固缩和核碎裂以及DNA片段化。综上所述，抑制UPK1A可抑制BTCC T24细胞的增殖并增强其凋亡，提示它是治疗这种疾病的潜在靶点。
