灯盏花乙素增强4T1乳腺癌细胞对顺铂敏感性的体内外研究

张琦; 包小波; 田冲冲

doi:10.13386/j.issn1002-0306.2023050219

灯盏花乙素增强4T1乳腺癌细胞对顺铂敏感性的体内外研究

Effect of Scutellarin on Promoting the Sensitivity of Breast Cancer 4T1 Cells to Cisplatin in Vitro and in Vivo

摘要

摘要: 本文旨在从体内外探究灯盏花乙素（Scutellarin，SCU）增强乳腺癌细胞4T1对顺铂（Cisplatin, CDDP）敏感性的作用及可能的分子机制。采用CCK-8实验、划痕实验、Transwell实验及流式细胞术体外分别探究SCU联合CDDP对4T1细胞增殖、迁移侵袭以及凋亡的影响；同时采用BALB/c小鼠皮下接种4T1细胞，建立荷瘤小鼠模型，在体观察SCU联合CDDP对肿瘤生长的作用；采用H&E染色方法对肿瘤组织进行形态学、坏死面积及微血管密度观察；分别采用实时荧光定量PCR与Western Blot检测肿瘤组织中凋亡因子转录水平及蛋白水平的表达。结果显示，SCU（200 μmol/L）联合CDDP（80 μmol/L）可极显著抑制4T1细胞的增殖（P<0.01）；SCU（200 μmol/L）联合CDDP（80 μmol/L）还能够极显著抑制4T1肿瘤细胞的迁移和侵袭能力（P<0.01），并且促进肿瘤细胞凋亡；60 mg/kg SCU联合3.0 mg/kg CDDP能够极显著抑制在体肿瘤4T1的生长（P<0.01）；且极显著抑制肿瘤微血管密度（P<0.01）；同时还能极显著促进肿瘤组织中促凋亡因子Caspase-3、Bax、Caspase-9、Cleaved-Caspase-3和Cleaved-Caspase-9的表达（P<0.01），显著抑制抗凋亡因子Bcl-2的表达（P<0.05）。综上所述，SCU通过增强4T1细胞对CDDP的敏感性，抑制肿瘤细胞功能且调控凋亡因子从而抑制肿瘤细胞的生长。

Abstract: To detect the effect of scutellarin (SCU) on promoting the sensitivity of breast cancer 4T1 cells to cisplatin (CDDP), and the molecular mechanism as well. CCK-8, scratch assay, Transwell assay and flow cytometry were employed to investigate the effects of SCU combined with CDDP on the proliferation, migration, invasion and apoptosis of 4T1 cells in vitro. Then, the 4T1 tumor-bearing mouse model was established to explore the effect of SCU combined with CDDP on tumor growth in vivo. The morphology, necrotic area and microvascular density of tumor tissues were observed by H&E staining. Real-time fluorescence quantitative PCR and Western Blot were used to detect the mRNA and proteins expression of apoptosis factors in tumor tissues. The results showed that SCU (200 μmol/L) combined with CDDP (80 μmol/L) significantly inhibited the in vitro proliferation of 4T1 cells (P<0.01). Moreover, the migration and invasion capacity of 4T1 cell were apparently reduced and the apoptosis of tumor cells were significantly promoted when treated with SCU (200 μmol/L) combined with CDDP (80 μmol/L) (P<0.01). Besides, the growth of 4T1 cells in vivo was remarkably slower after administration of 60 mg/kg SCU combined with 3.0 mg/kg CDDP (P<0.01). Further H&E semi-quantitative results revealed the microvascular density of 4T1 tumor tissues was significantly decreased (P<0.01). What’s more, the combination of SCU and CDDP significantly promoted the expression of pro-apoptotic factors Caspase-3, Bax, Caspase-9, Cleaved-Caspase-3 and Cleaved-Caspase-9 (P<0.01) and inhibited the expression of anti-apoptotic factor Bcl-2 in tumor tissues (P<0.05). In conclusion, SCU combined with CDDP could inhibit the function of tumor cells by enhancing the sensitivity of 4T1 cells to CDDP, and also regulate the expression of apoptosis factors, thus inhibiting the growth of tumor.