| 许卫星,张薇,陈姣敏,等.苦参碱调节 Hippo-Yes相关蛋白信号通路对人非霍奇金淋巴瘤 Raji细胞增殖和凋亡的影响[J].安徽医药,2025,29(11):2165-2170. |
| 苦参碱调节 Hippo-Yes相关蛋白信号通路对人非霍奇金淋巴瘤 Raji细胞增殖和凋亡的影响 |
| Impacts of matrine on the proliferation and apoptosis of human non-Hodgkin's lymphoma Raji cells by regulating Hippo-YAP signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2025.11.009 |
| 中文关键词: 苦参碱 淋巴瘤，非霍奇金 Burkitt淋巴瘤细胞(Raji细胞) 增殖 凋亡 Hippo-Yes相关蛋白信号通路 |
| 英文关键词: Matrine Lymphoma, non-Hodgkin Burkitt lymphoma cells (Raji cells) Proliferation Apoptosis Hippo-Yes associated protein signaling pathway |
| 基金项目:河北省中医药管理局科研计划项目( 2023454) |
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| 中文摘要: |
| 目的探究苦参碱调节 Hippo-Yes相关蛋白( YAP)信号通路对人非霍奇金淋巴瘤( NHL)Raji细胞增殖和凋亡的影响。方法 2023年 1—7月，以不同浓度苦参碱( 0、0.25、0.50、1.00、2.00、4.00、8.00 mmol/L)处理 Raji细胞，细胞计数试剂盒( CKK-8)法检测细胞增殖情况。后续根据 Raji细胞在 48 h的半抑制浓度( IC50)，将 Raji细胞分为对照组(正常培养，不作处理)，苦参碱低、中、高剂量组(分别以 1、2、4 mmol/L苦参碱处理)苦参碱高剂量 +Hippo信号通路抑制剂( XMU-MP-1)组(以 4 mmol/L苦参碱和 1 μmol/L XMU-MP-1处理)。分别以集落形成试验，检测各组细胞集落形成数，流式细胞术检测细胞周期和凋亡，蛋白质印迹法检测大肿瘤抑制因子 1(LATS1)、磷酸化 LATS1(p-LATS1)、 YAP、具有 PDZ基序的转录共激活因子( TAZ)、细胞周期蛋白 D1(cyclin D1)、核增殖抗原( PCNA)、 B细胞淋巴瘤 -2(Bcl-2)、 Bcl-2相关 X蛋白( Bax)蛋白表达。结果与空白对照( 0 mmol/L苦参碱浓度)组［( 0±0)%］相比， 0.25［( 9.48±1.05)%、(10.25±1.36)%、(12.57±1.46)%］、 0.50［( 16.37±1.82)%、(19.62±2.05)%、(21.95±2.26)%］、 1.00［( 35.72±4.68)%、(37.58±3.92)%、(38.42±3.51)%］、 2.00［( 49.06±5.29)%、(51.49±5.36)%、(54.86±5.93)%］、 4.00［( 59.37±5.75)%、(68.62±6.85)%、(73.82±7.57)%］、 8.00 mmol/L［( 80.59±8.57)%、(82.73±8.84)%、(83.05±8.74)%］苦参碱能够显著随浓度增加抑制 Raji细胞 24、48和 72 h增殖活性( P<0.05)。与对照组相比，苦参碱低、中、高剂量组细胞集落形成数［( 96.28±10.06)个、(73.26±7.38)个、(48.35±4.68)个比( 136.85±14.61)个］显著降低( P<0.05)，细胞凋亡率［( 13.74±1.38)%、(21.62±2.57)%、(28.15±3.24)%比( 4.82±0.53)%］显著增加( P<0.05)S期细胞比例、 cyclin D1、PCNA、Bcl-2、 YAP、TAZ蛋白表达显著降低( P<0.05)G1期细胞比例、 Bax、p-LATS1/LATS1表达显著增加(P<0.05)；与苦参碱高剂量组相比，苦参碱高剂量 +XMU-MP-1组细胞集落形成数、 S期细胞比例、 cyclin D1、PCNA、Bcl-2、YAP、TAZ蛋白表达显著增加(P<0.05)G1期细胞比例、细胞凋亡率、 Bax、p-LATS1/LATS1表达显著下降( P<0.05)。结论苦参碱可能通过调节 Hippo-YAP信号通路,抑制 NHL Raji细胞增殖，促进细胞周期阻滞和凋亡。 |
| 英文摘要: |
| Objective To investigate the impacts of matrine on the proliferation and apoptosis of human non-Hodgkin's lymphoma (NHL) Raji cells by regulating the Hippo-Yes associated protein (YAP) signaling pathway.Methods From January to July 2023, Rajicells were treated with different concentrations of matrine (0, 0.25, 0.50, 1.00, 2.00, 4.00, 8.00 mmol/L), and cell proliferation was de-tected using the cell counting kit-8 (CKK-8) method. Subsequently, based on the half maximal inhibitory concentration (IC50) of Rajicells at 48 h, the cells were separated into control group (normal culture, no treatment), low, medium, and high dose matrine groups(treated with 1, 2, 4 mmol/L matrine respectively), and high dose matrine+Hippo signaling pathway inhibitors (XMU-MP-1) group (treat-ed with 4 mmol/L matrine and 1 μmol/L XMU-MP-1), the colony formation test was applied to detect the number of colony formation ofcells in each group, flow cytometry was applied to detect cell cycle and apoptosis, Western blotting was applied to detect the expressionof large tumor suppressor gene 1 (LATS1), phosphorylated LATS1 (p-LATS1), YAP, transcriptional coactivator with PDZ-binding motif (TAZ), cyclin D1, proliferating cell nuclear antigen (PCNA), B-cell lymphoma factor 2 (Bcl-2) and Bcl-2 related protein (Bax) proteins. Results Compared with the blank control (0 mmol/L matrine concentration) group [(0±0)% ], 0.25 [(9.48±1.05)% , (10.25±1.36)% ,(12.57±1.46)%], 0.50 [(16.37±1.82)%, (19.62±2.05)%, (21.95±2.26)%], 1.00 [(35.72±4.68)%, (37.58±3.92)%, (38.42±3.51)%], 2.00[(49.06±5.29)% , (51.49±5.36)% , (54.86±5.93)% ], 4.00 [(59.37±5.75)% , (68.62±6.85)% , (73.82±7.57)% ], 8.00 mmol/L [(80.59±8.57)%, (82.73±8.84)%, (83.05±8.74)%] matrine greatly inhibited the proliferation activity of Raji cells at 24, 48 and 72 h with increas-ing concentration (P<0.05). Compared with the control group, the number of cell colonies formed (96.28±10.06, 73.26±7.38, 48.35±4.68 vs. 136.85±14.61) were greatly reduced in low, medium, and high dose matrine groups (P<0.05), the apoptosis rate [(13.74± 1.38)%, (21.62±2.57)%, (28.15±3.24)% vs. (4.82±0.53)%] were greatly increased (P<0.05), the proportion of S phase cells, and the ex-pression of cyclin D1, PCNA, Bcl-2, YAP, and TAZ proteins were greatly reduced (P<0.05), the proportion of G1 phase cells, and the expression of Bax and p-LATS1/LATS1 were greatly increased (P<0.05); compared with the high dose matrine group, the number of cellcolonies formed, the proportion of S phase cells, and the expression of cyclin D1, PCNA, Bcl-2, YAP, and TAZ proteins were greatly in-creased the high dose matrine+XMU-MP-1 group (P<0.05), the proportion of G1 phase cells, apoptosis rate, the expression of Bax and p-LATS1/LATS1 were greatly reduced (P<0.05).Conclusion Matrine may inhibit proliferation of NHL Raji cells, and promote cell cy-cle arrest and apoptosis by regulating the Hippo-YAP signaling pathway. |
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