Department of Gynecology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Gynecology, People's Hospital of Tang County, Baoding, Hebei, China.
Histol Histopathol. 2021 May;36(5):535-545. doi: 10.14670/HH-18-310. Epub 2021 Feb 11.
To investigate the effect of the downregulation of FXYD domain-containing ion transport regulator 5 (FXYD5) on the cisplatin resistance (CisR) of epithelial ovarian cancer (EOC) cells.
A2780-CisR and SKOV3-CisR cells were obtained through repeated administrations of different cisplatin concentrations, and the half-maximal inhibition concentration (IC50) was calculated by MTT assays. After transfection with FXYD5 siRNA-1 and FXYD5 siRNA-2, the IC50 values of the A2780-CisR and SKOV3-CisR cells were also detected by the MTT method. Cell proliferation, migration, invasion and apoptosis were evaluated through 5-ethynyl-2'-deoxyuridine (EdU) DNA synthesis, wound healing, Transwell invasion and Annexin-V-FITC/PI dual-staining assays, respectively. qRT-PCR and Western blotting were conducted to detect mRNA and protein expression.
Compared with the sensitive parental cells, the A2780-CisR and SKOV3-CisR cells had increased IC50 and FXYD5 expression. FXYD5 siRNA reduced the IC50 value of cisplatin in the A2780-CisR and SKOV3-CisR cells and decreased the expression of ABCG2 (BCRP) and ABCB1 (MDR1). In addition, FXYD5 inhibition reduced the invasion and migration of the A2780-CisR and SKOV3-CisR cells, with upregulation of E-cadherin and downregulation of Snail and Vimentin. Both FXYD5 siRNA-1 and FXYD5 siRNA-2 inhibited the proliferation and promoted the apoptosis of the A2780-CisR and SKOV3-CisR cells with reduced Ki-67 and increased caspase-3.
FXYD5 downregulation may reduce the invasion, migration and EMT formation of EOC cells to increase their sensitivity to cisplatin chemotherapy by inhibiting cell proliferation and promoting cell apoptosis.
研究 FXYD 结构域含离子转运调节剂 5（FXYD5）下调对上皮性卵巢癌（EOC）细胞顺铂耐药（CisR）的影响。
通过反复给予不同浓度顺铂，获得 A2780-CisR 和 SKOV3-CisR 细胞，并通过 MTT 测定计算半抑制浓度（IC50）。用 FXYD5 siRNA-1 和 FXYD5 siRNA-2 转染后，通过 MTT 法检测 A2780-CisR 和 SKOV3-CisR 细胞的 IC50 值。通过 5-乙炔基-2'-脱氧尿苷（EdU）DNA 合成、划痕愈合、Transwell 侵袭和 Annexin-V-FITC/PI 双重染色试验分别评估细胞增殖、迁移、侵袭和凋亡。通过 qRT-PCR 和 Western blot 检测 mRNA 和蛋白表达。
与敏感亲本细胞相比，A2780-CisR 和 SKOV3-CisR 细胞的 IC50 和 FXYD5 表达增加。FXYD5 siRNA 降低了 A2780-CisR 和 SKOV3-CisR 细胞中顺铂的 IC50 值，并降低了 ABCG2（BCRP）和 ABCB1（MDR1）的表达。此外，FXYD5 抑制降低了 A2780-CisR 和 SKOV3-CisR 细胞的侵袭和迁移，上调了 E-钙黏蛋白，下调了 Snail 和波形蛋白。FXYD5 siRNA-1 和 FXYD5 siRNA-2 均抑制了 A2780-CisR 和 SKOV3-CisR 细胞的增殖，促进了细胞凋亡，Ki-67 减少，caspase-3 增加。
FXYD5 下调可能通过抑制细胞增殖和促进细胞凋亡，减少 EOC 细胞的侵袭、迁移和 EMT 形成，从而提高其对顺铂化疗的敏感性。
