| 引用本文: | 徐加杰,葛明华,郑国湾,王林红,郭海魏,郑传铭.乌骨藤中C21甾体苷诱导SACC-83和SACC-LM细胞凋亡的作用及其机制研究[J].中国现代应用药学,2019,36(10):1205-1211. |
| XU Jiajie,GE Minghua,ZHENG Guowan,WANG Linhong,GUO Haiwei,ZHENG Chuanming.Apoptosis Induced by the C21 Sterols in Marsdeniae Tenacissimae Caulis and Its Molecule Mechanism of Action in SACC-83 and SACC-LM Cells[J].Chin J Mod Appl Pharm(中国现代应用药学),2019,36(10):1205-1211. |
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| 摘要: |
| 目的 研究乌骨藤提取物C21甾体苷对唾液腺腺样囊性癌（salivary adenoid cystic carcinomacv，SACC）低侵袭细胞株（salivary adenoid cystic carcinoma-83，SACC-83）和肺高转移细胞株（SACC-LM）增殖抑制和诱导凋亡的作用及其机制。方法 用不同浓度（5，10，20，40，60，80，100 μmol·L-1） C21甾体苷处理SACC-83和SACC-LM细胞48 h后，MTT法检测细胞活力，并计算药物的IC20和IC50；细胞克隆形成实验检测细胞增殖能力；流式细胞术检测SACC-83及SACC-LM细胞凋亡情况；实时荧光定量PCR和Western blot检测SACC-83和SACC-LM细胞Bcl-2、Bax、caspase 3的mRNA和蛋白的表达。结果 不同浓度的C21甾体苷降低SACC-83和SACC-LM细胞活力，抑制细胞增殖，并且作用于SACC-83细胞C21甾体苷的IC20浓度为7.49 μmol·L-1，IC50浓度为38.34 μmol·L-1；作用于SACC-LM细胞的C21甾体苷IC20浓度为9.30 μmol·L-1，IC50浓度为46.04 μmol·L-1；细胞克隆集落形成明显减少。C21甾体苷IC20浓度分别促进SACC-83及SACC-LM细胞凋亡，且随着给药时间延长，凋亡率增加，具有显著性差异（P<0.05，P<0.01）。经7.49，9.30 μmol·L-1 C21甾体苷分别处理SACC-83及SACC-LM细胞后，Bcl-2的mRNA及蛋白水平显著降低（P<0.01），而Bax、Caspase 3的mRNA和蛋白水平显著升高（P<0.05）。结论 乌骨藤C21甾体苷抑制SACC-83及SACC-LM细胞增殖、促进凋亡，其作用机制可能与调控Bcl-2、Bax和Caspase 3表达有关。 |
| 关键词: 乌骨藤 C21 甾体苷 SACC-83细胞 SACC-LM细胞 细胞增殖 细胞凋亡 |
| DOI：10.13748/j.cnki.issn1007-7693.2019.10.007 |
| 分类号:R285.5 |
| 基金项目:浙江省中医药科技计划项目（2018ZQ010） |
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| Apoptosis Induced by the C21 Sterols in Marsdeniae Tenacissimae Caulis and Its Molecule Mechanism of Action in SACC-83 and SACC-LM Cells |
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XU Jiajie, GE Minghua, ZHENG Guowan, WANG Linhong, GUO Haiwei, ZHENG Chuanming
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Zhejiang Provincial People's Hospital, Hangzhou 310014, China
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| Abstract: |
| OBJECTIVE To evaluate the effects and mechanism of C21 sterols in Marsdeniae Tenacissimae Caulis extract on inhibition of proliferation and induction of apoptosis of low invasion and lung hypermetastasis in salivary adenoid cystic carcinomacv cell line SACC-83 and SACC-LM, respectively. METHODS SACC-83 and SACC-LM cells were treated with C21 sterols at different doses (5, 10, 20, 40, 60, 80, 100 μmol·L-1) for 48 h, then the cell viability was measured by MTT assay, and calculated the value of IC20 and IC50 for drug. The capacity of clone formation of cells were detected by clone formation assay. The apoptotic of SACC-83 and SACC-LM cells were determined by flow cytometry. The expression levels of Bcl-2, Bax and caspase-3 mRNA and proteins in SACC-83 and SACC-LM cells were examined by q-PCR and Western blot method. RESULTS C21 sterols at different doses decreased the viability and inhibited proliferation of SACC-83 and SACC-LM cells, and the concentration of IC20 was 7.49 μmol·L-1, and IC50 was 38.34 μmol·L-1 for SACC-83 cells. In addition, the concentration of IC20 was 9.30 μmol·L-1, and IC50 was 46.04 μmol·L-1 for SACC-LM cells. The apoptotic rate of SACC-83 and SACC-LM cells under concentration of IC20 with C21 sterols was significantly increased(P<0.05, P<0.01). The q-PCR and Western blot results showed that the expression level of Bcl-2 in the SACC-83 and SACC-LM cells was decreased significantly(P<0.01), and the expression level of Bax and Caspase-3 were increased significantly (P<0.05). CONCLUSION C21 sterols in Marsdeniae Tenacissimae Caulis extract can depress the proliferation and promote apoptosis of SACC-83 and SACC-LM cells, which mechanism may be related to decreasing the Bcl-2 level, and increasing the Bax and Caspase-3 level. |
| Key words: Marsdeniae Tenacissimae Caulis C21 sterols SACC-83 cell SACC-LM cell cell proliferation cell apoptosis |
