| 李霞,李慧,许聪聪,等.肉灵芝提取物调控长链非编码 RNA BRCA1相邻基因 2/双特异性磷酸酶 4/细胞外信号调节激酶通路对肝癌细胞 Hep3B增殖及凋亡的影响[J].安徽医药,2022,26(11):2208-2212. |
| 肉灵芝提取物调控长链非编码 RNA BRCA1相邻基因 2/双特异性磷酸酶 4/细胞外信号调节激酶通路对肝癌细胞 Hep3B增殖及凋亡的影响 |
| Effects of Ganoderma lucidum extract on the proliferation and apoptosis of Hep3B hepatocellular carcinoma cells by regulating the lncRNA NBR2/DUSP4/ERK pathway |
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| DOI：10.3969/j.issn.1009-6469.2022.11.020 |
| 中文关键词: 肉灵芝提取物 长链非编码 RNA BRCA1相邻基因 2 DUSP4/ERK 肝癌 |
| 英文关键词: Ganoderma lucidum extract LncRNA NBR2 DUSP4/ERK signaling pathway Liver cancer |
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| 中文摘要: |
| 目的探讨肉灵芝提取物是否可通过长链非编码 RNA BRCA1相邻基因 2(LncRNA NBR2)/双特异性磷酸酶 4/细胞外信号调节激酶( DUSP4/ERK)通路而影响肝癌 HepB细胞增殖及凋亡。方法体外培养肝癌 HepB细胞，分为 1 mg/L肉灵芝提取物组、 2 mg/L肉灵芝提取物组、 4 mg/L肉灵芝提取物组、 si-NC组、 si-LncRNA NBR2组、 pcDNA-NC组、 pcDNA-LncRNA NBR2组、肉灵芝提取物 +si-NC组、肉灵芝提取物 +si-LncRNA NBR2组；采用 MTT检测肝癌 HepB细胞增殖；流式细胞术检测肝癌 HepB细胞凋亡； RT-qPCR检测 LncRNA NBR2的表达量；蛋白质印迹法( Western blotting)检测增殖细胞核抗原(PCNA)、活化的含半胱氨酸的天冬氨酸蛋白水解酶 3(Cleaved-caspase-3)、 DUSP4、p-ERK蛋白表达量。结果肉灵芝提取物可明显降低肝癌 HepB细胞增殖率与 PCNA、DUSP4、p-ERK蛋白水平( P<0.05)，提高肝癌 HepB细胞凋亡率、 Cleaved-caspase-3蛋白水平及 Ln. cRNA NBR2的表达水平( P<0.05)；转染 si-LncRNA NBR2后可明显提高肝癌 HepB细胞增殖率及 PCNA、DUSP4、p-ERK蛋白水平( P<0.05)降低凋亡率及 Cleaved-caspase-3蛋白水平( P<0.05)而转染 pcDNA-NBR2可明显减弱转染 si-LncRNA NBR2对肝癌 HepB细胞增，殖及凋亡的作用；与肉灵芝提取物 +si-NC组比较，，肉灵芝提取物 +si-LncRNA NBR2组肝癌 HepB细胞增殖率及 PCNA、DUSP4、p-ERK蛋白水平升高( P<0.05)，凋亡率及 Cleaved-caspase-3蛋白水平降低( P<0.05)。结论肉灵芝提取物可通过上调 LncRNA NBR2的表达而抑制 DUSP4/ERK信号通路的活化从而抑制肝癌 HepB细胞增殖及诱导细胞凋亡。 |
| 英文摘要: |
| Objective To investigate whether Ganoderma lucidum extract can affect the proliferation and apoptosis of Hep3B livercancer cells through the long noncoding RNA BRCA1 adjacent gene 2 (lncRNA NBR2)/dual specificity phosphatase 4/extracellular sig.nal-regulated kinase (DUSP4/ERK) pathway.Methods Hep3B cells were cultured in vitro and divided into the 1 mg/L Ganoderma lu.cidum extract group, 2 mg/L Ganoderma lucidum extract group, 4 mg/L Ganoderma lucidum extract group, si-NC group, si-lncRNA NBR2 group, pcDNA-NC group, pcDNA-lncRNA NBR2 group, Ganoderma extract+si-NC group, and Ganoderma extract+si-lncRNA NBR2 group. MTT was used to detect the proliferation of Hep3B cells, and flow cytometry was used to detect the apoptosis of Hep3Bcells. RT.qPCR was used to detect the expression of lncRNA NBR2; Western blot was used to detect proliferating cell nuclear antigen(PCNA), activated cysteine-containing caspase-3 (Cleaved-caspase-3), DUSP4, and p-ERK protein expression.Results Ganoderma lu. cidum extract can significantly reduce the proliferation rate of Hep3B cells and the protein levels of PCNA, DUSP4, and p-ERK (P < 0.05) and increased the apoptosis rate and the expression levels of cleaved caspase-3 and lncRNA NBR2 in Hep3B cells (P < 0.05). Transfection of si-lncRNA NBR2significantly increased the proliferation rate and the protein levels of PCNA, DUSP4, and p-ERK (P < 0.05) and decreased the apoptosis rate and the protein level of cleaved-caspase-3(P < 0.05). Transfection of pcDNA-lncRNA NBR2 sig. nificantly attenuated the effect of si-lncRNA NBR2 transfection on the proliferation and apoptosis of Hep3B cells. Compared with the Ganoderma lucidum extract+si-NC group, the cell proliferation rate and the protein levels of PCNA, DUSP4, and p-ERK in the Gano. derma lucidum extract+si-LncRNA NBR2 group increased (P<0.05), while apoptosis and the protein level of cleaved-caspase-3 were re. duced (P < 0.05).Conclusion Ganoderma lucidum extract significantly reduced the activation of the DUSP4/ERK signaling pathwayby upregulating the expression of LncRNA NBR2, thereby inhibiting the proliferation and increased the apoptosis of Hep3B cells. |
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