紫草素对卵巢癌细胞株SKOV3放疗敏感性的影响及相关机制研究
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摘要
目的:研究紫草素对人卵巢癌细胞株SKOV-3的放射增敏作用,并探讨其可能的作用机制。方法:MTT法检测不同浓度(0、5、10、20、40、80和120 mg/L)紫草素对SKOV-3细胞活力的影响;克隆形成实验检测8 mg/L紫草素联合不同剂量(0、2、4、6和8 Gy)X射线照射对SKOV-3细胞增殖能力的影响。将SKOV-3细胞分为4组:对照组、紫草素组(8 mg/L紫草素处理)、8 Gy组(8 Gy X射线照射处理)和紫草素+8 Gy组(给予8 mg/L紫草素处理48 h后,再进行8 Gy X射线照射处理)。PI染色及流式细胞术检测各组SKOV-3细胞周期的变化;Western blot检测各组SKOV-3细胞p-PI3K和p-AKT的蛋白表达水平。结果:在0～80 mg/L浓度范围内,紫草素浓度依赖性抑制SKOV-3细胞活力(P<0.05),IC_(50)为(38.54±0.57) mg/L;紫草素联合放疗处理后的SKOV-3细胞的存活曲线较单独放疗左移,放射增敏比为1.45±0.05。与8 Gy组相比,紫草素+8 Gy组G_2/M期细胞所占比例下降(P<0.05),p-PI3K和p-AKT蛋白水平均显著降低(P<0.05)。结论:紫草素具有提高人卵巢癌细胞株SKOV-3放射敏感性的作用,其作用机制可能与紫草素缓解X射线引起的G_2/M期阻滞及抑制PI3K/AKT信号通路有关。
AIM: To investigate the radiosensitizing effect of shikonin on the human ovarian cancer SKOV-3 cells and the underlying mechanism. METHODS: The viability of SKOV-3 cells after treated with shikonin at different concentrations(0, 5, 10, 20, 40, 80 and 120 mg/L) was measured by MTT assay. The proliferation ability of SKOV-3 cells after treated with different doses(0, 2, 4, 6 and 8 Gy) of X-ray radiotherapy was tested by colony formation assay. The SKOV-3 cells were divided into 4 groups: control group, shikonin group(8 mg/L shikonin treatment), 8 Gy group(8 Gy X-ray treatment) and shikonin+8 Gy group(8 mg/L shikonin treatment for 48 h, followed by 8 Gy X-ray treatment). The cell cycle was examined by PI staining and flow cytometry, and the phosphorylation levels of PI3 K and AKT were analyzed by Western blot. RESULTS: In the range of 0～80 mg/L, shikonin decreased the viability of SKOV-3 cells in a concentration-dependent manner(P<0.05). The value of IC_(50) was(38.54±0.57) mg/L. Compared with radiotherapy alone, the survival curve was markedly shifted to the left after shikonin treatment combined with radiotherapy(P<0.05). The value of radiotherapy sensitization enhancement ratio was 1.45±0.05. Moreover, compared with 8 Gy alone group, the percentage of G_2/M phase and the phosphorylation levels of PI3 K and AKT were decreased in shikonin+8 Gy group(P<0.05). CONCLUSION: Shikonin increases the radiosensitivity of SKOV-3 cells, and the mechanism may be related to attenuation of radiation-induced G_2/M phase arrest and inhibition of PI3 K/AKT signaling pathway.
AIM: To investigate the radiosensitizing effect of shikonin on the human ovarian cancer SKOV-3 cells and the underlying mechanism. METHODS: The viability of SKOV-3 cells after treated with shikonin at different concentrations(0, 5, 10, 20, 40, 80 and 120 mg/L) was measured by MTT assay. The proliferation ability of SKOV-3 cells after treated with different doses(0, 2, 4, 6 and 8 Gy) of X-ray radiotherapy was tested by colony formation assay. The SKOV-3 cells were divided into 4 groups: control group, shikonin group(8 mg/L shikonin treatment), 8 Gy group(8 Gy X-ray treatment) and shikonin+8 Gy group(8 mg/L shikonin treatment for 48 h, followed by 8 Gy X-ray treatment). The cell cycle was examined by PI staining and flow cytometry, and the phosphorylation levels of PI3 K and AKT were analyzed by Western blot. RESULTS: In the range of 0～80 mg/L, shikonin decreased the viability of SKOV-3 cells in a concentration-dependent manner(P<0.05). The value of IC_(50) was(38.54±0.57) mg/L. Compared with radiotherapy alone, the survival curve was markedly shifted to the left after shikonin treatment combined with radiotherapy(P<0.05). The value of radiotherapy sensitization enhancement ratio was 1.45±0.05. Moreover, compared with 8 Gy alone group, the percentage of G_2/M phase and the phosphorylation levels of PI3 K and AKT were decreased in shikonin+8 Gy group(P<0.05). CONCLUSION: Shikonin increases the radiosensitivity of SKOV-3 cells, and the mechanism may be related to attenuation of radiation-induced G_2/M phase arrest and inhibition of PI3 K/AKT signaling pathway.
引文
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[10] Hufnagl A, Herr L, Friedrich T, et al. The link between cell-cycle dependent radiosensitivity and repair pathways: a model based on the local, sister-chromatid conformation dependent switch between NHEJ and HR[J]. DNA Repair (Amst), 2015, 27:28-39.
[11] Wang L, Xia Y, Chen T, et al. Sanyang Xuedai enhances the radiosensitivity of human non-small cell lung cancer cells via increasing iNOS/NO production[J]. Biomed Pharmacother, 2018, 102:618-625.
[12] Luo Y, Chen X, Luo L, et al. [6]-Gingerol enhances the radiosensitivity of gastric cancer via G2/M phase arrest and apoptosis induction[J]. Oncol Rep, 2018, 39(5):2252-2260.
[13] Pan S, Sun Y, Sui D, et al. Lobaplatin promotes radiosensitivity, induces apoptosis, attenuates cancer stemness and inhibits proliferation through PI3K/AKT pathway in esophageal squamous cell carcinoma[J]. Biomed Pharmacother, 2018, 102:567-574.
[14] Yang XX, Ma M, Sang MX, et al. BMI-1 suppression increases the radiosensitivity of oesophageal carcinoma via the PI3K/Akt signaling pathway[J]. Oncol Rep, 2018,39(2):667-678.
[2] 田红, 于鹏, 吴小茗, 等. 卵巢癌的治疗药物研究进展[J].现代药物与临床, 2015, 30(1):103-107.
[3] Yan Y, Furumura M, Gouya T, et al. Shikonin promotes skin cell proliferation and inhibits nuclear factor-κB translocation via proteasome inhibition in vitro[J]. Chin Med J (Engl), 2015, 128(16):2228-2233.
[4] Li Y, Lu H, Gu Y, et al. Enhancement of NK cells proliferation and function by Shikonin[J]. Immunopharmacol Immunotoxicol, 2017, 39(3):124-130.
[5] 陈菊英, 刘朝纯, 曾智, 等. 紫草素通过PI3K/Akt通路促进人乳腺癌MCF-7细胞自噬[J]. 中国药理学通报, 2013, 29(2): 194-198.
[6] 于海荣, 苗浩, 庞冲, 等. 紫草素对人宫颈癌Hela细胞增殖的影响及其作用机制研究[J]. 中国生化药物杂志, 2015, 35(4):16-18.
[7] Shan ZL, Zhong L, Xiao CL, et al. Shikonin suppresses proliferation and induces apoptosis in human leukemia NB4 cells through modulation of MAPKs and c-Myc[J].Mol Med Rep, 2017, 16(3):3055-3060.
[8] 王汝兴, 鲁艳杰, 周健, 等. 紫草素对人卵巢癌细胞SKOV3细胞增殖和凋亡的影响[J].中药药理与临床, 2016, 32(2):76-79.
[9] Im MM, Flanagan SA, Ackroyd JJ, et al. Drug metabolism and homologous recombination repair in radiosensitization with gemcitabine[J].Radiat Res, 2015, 183(1):114-123.
[10] Hufnagl A, Herr L, Friedrich T, et al. The link between cell-cycle dependent radiosensitivity and repair pathways: a model based on the local, sister-chromatid conformation dependent switch between NHEJ and HR[J]. DNA Repair (Amst), 2015, 27:28-39.
[11] Wang L, Xia Y, Chen T, et al. Sanyang Xuedai enhances the radiosensitivity of human non-small cell lung cancer cells via increasing iNOS/NO production[J]. Biomed Pharmacother, 2018, 102:618-625.
[12] Luo Y, Chen X, Luo L, et al. [6]-Gingerol enhances the radiosensitivity of gastric cancer via G2/M phase arrest and apoptosis induction[J]. Oncol Rep, 2018, 39(5):2252-2260.
[13] Pan S, Sun Y, Sui D, et al. Lobaplatin promotes radiosensitivity, induces apoptosis, attenuates cancer stemness and inhibits proliferation through PI3K/AKT pathway in esophageal squamous cell carcinoma[J]. Biomed Pharmacother, 2018, 102:567-574.
[14] Yang XX, Ma M, Sang MX, et al. BMI-1 suppression increases the radiosensitivity of oesophageal carcinoma via the PI3K/Akt signaling pathway[J]. Oncol Rep, 2018,39(2):667-678.