| 李晨,李占恩,苏宏伟,等.不同浓度苍术素通过 RhoA/ROCK信号通路对膀胱癌细胞生物学行为的影响[J].安徽医药,2026,30(11):2185-2189. |
| 不同浓度苍术素通过 RhoA/ROCK信号通路对膀胱癌细胞生物学行为的影响 |
| Study on apoptosis and autophagy of bladder cancer cells induced by regulation of RhoA/ROCK signaling pathway by atractylotin |
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| DOI：10.3969/j.issn.1009-6469.2026.11.012 |
| 中文关键词: 苍术 膀胱癌 RhoA/ROCK信号通路 凋亡 自噬 |
| 英文关键词: Atractylodes rhizome Bladder cancer RhoA/ROCK signaling pathway Apoptosis Autophagy |
| 基金项目:张家口市科学技术局重点研发技术项目( 2221129D) |
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| 中文摘要: |
| 目的分析不同浓度苍术素通过 RhoA/Rho激酶( ROCK)信号通路对膀胱癌细胞的凋亡及自噬作用。方法 2022年 9月至 2023年 9月将膀胱癌 T24细胞采用随机数字表法分组、建模，分为对照组、低浓度苍术素组( L-苍术素组)、中浓度苍术素组( M-苍术素组)、高浓度苍术素组( H-苍术素组)及高浓度苍术素 +U46619组( H-苍术素 +U46619组)。以细胞计数试剂盒 -8(CCK-8)法测定癌细胞增殖活性，包括 450 nm处的吸光度［D(λ)450 nm］及细胞抑制率；以流式细胞术( FCM)测定细胞凋亡率；蛋白质印迹法检测自噬相关蛋白及 RhoA/ROCK信号通路相关蛋白，前者包括 LC3-Ⅰ、LC3-Ⅱ、Beclin-1，后者包括 RhoA、ROCK1蛋白。结果培养 48 h及 72 h时， L、M、H-苍术素组 D(λ)450 nm值均低于对照组，细胞抑制率、凋亡率均高于对照组( P<0.05)，且随着浓度增加，变化越显著( P<0.05)。与对照组比较，各浓度苍术素组 LC3-Ⅱ/LC3-Ⅰ、Beclin-1均更高( P<0.05)，但 RhoA、 ROCK1含量则更低( P<0.05)；借助 U46619激活 RhoA/ROCK信号通路后， H-苍术素 +U46619组的 D(λ)450 nm值均高于 H-苍术素组［48 h:(0.96±0.08)比( 0.68±0.06)； 72 h:(1.00±0.40)比( 0.86±0.27)］，且细胞抑制率［48 h:(2.39±0.14)％， 72 h:(35.76±3.04)％］、凋亡率［48 h:(15.47±2.00)％， 72 h:(18.86±1.58)％］及 LC3-Ⅱ/LC3-Ⅰ比值( 1.39±0.13)、 Beclin-1(1.42±0.10)/GAPDH蛋白、 RhoA(1.02±0.09)/GAPDH、ROCK1蛋白含量( 1.22±0.07)/GAPDH均与 H-苍术素组差异有统计学意义( P<0.05)。结论苍术素能显著调节膀胱癌细胞的增殖活性，对其自噬能力亦有调节效果，应用剂量越大、上述效应越显著，可能与苍术素抑制了 RhoA/ROCK信号通路有关。 |
| 英文摘要: |
| Objective To analyze the effects of different concentrations of Atractylodes on apoptosis and autophagy in bladder cancercells via the RhoA/ROCK signaling pathway.Methods From September 2022 to September 2023, T24 cells were categorized into five groups: control group, low-concentration Atractylodes group (L-atractylodes), medium-concentration Atractylodes group (M-atracty-lodes), high-concentration Atractylodes group (H-atractylodes), and high-concentration Atractylodes + U46619 group (H-atractylodes + U46619). The proliferative activity of cancer cells was assessed using the CCK-8 method, measuring absorbance [D(λ)450 nm value] andcell inhibition rate. Apoptosis rates were evaluated by flow cytometry (FCM). Western blotting was employed to detect proteins relatedto autophagy, including LC3-Ⅰ , LC3-Ⅱ , Beclin-1, as well as proteins associated with the RhoA/ROCK signaling pathway such as RhoA and ROCK1.Results At 48 h and 72 h, D(λ)450 nm values of L-,M-, and H-atractylodes groups were significantly lower than thoseof the control group; correspondingly, both cell inhibition rates and apoptosis rates were higher compared to controls (P<0.05), with more pronounced changes observed at increased concentrations (P<0.05). Compared to the control group, levels of LC3-Ⅱ/LC3-Ⅰ ratio and Beclin-1 in Atractylodes groups were elevated (P<0.05), while contents of RhoA and ROCK1 decreased significantly (P<0.05). Fol-lowing activation of the RhoA/ROCK signaling pathway with U46619, D(λ)450 nm values for H-atractylosin + U46619 exceeded those for H-atractylosin [48 h: (0.96±0.08) vs. (0.68±0.06); 72 h: (1.00±0.40) vs. (0.86±0.27)]. The cell inhibition rate [48 h: (2.39±0.14)%, 72 h:(35.76±3.04)%], apoptosis rate [48 h: (15.47±2.00)%, 72 h: (18.86±1.58)%], LC3-Ⅱ/LC3-Ⅰ ratio (1.39±0.13), Beclin-1 (1.42±0.10)/ GAPDH protein, RhoA (1.02±0.09)/GAPDH protein, and ROCK1 protein content (1.22±0.07)/GAPDH protein] in H-atractylosin + U46619 group were all statistically significantly different from those in the H-atractyloidsin group (P<0.05).Conclusion The rhizoma atractyloidis component can significantly modulate proliferation activity in bladder cancer cells; its influence on autophagic capacity isalso notable-greater doses yield more substantial effects-potentially through inhibition of the RhoA/ROCK signaling pathway. |
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