黑果枸杞花色苷Pt3G对前列腺癌LNCaP和PC-3细胞增殖与凋亡的影响  

Effects of Lycium ruthenicum Murray anthocyanin Pt3G on the proliferation and apoptosis of prostate cancer LNCaP and PC-3 cells

作　　者：李施莹 李占龙 米佳 禄璐 闫亚美 曹有龙 冉林武 杨文 Shiying Li;Zhanlong Li;Jia Mi;Lu Lu;Yamei Yan;Youlong Cao;Linwu Ran;Wen Yang(School of Public Health and Management,Ningxia Medical University,Yinchuan 750004,China;Goji Berry Research Institute,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002,China;Laboratory Animal Center,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院,银川750004 [2]宁夏农林科学院枸杞科学研究所,银川750002 [3]宁夏医科大学实验动物中心,银川750004

出　　处：《科学通报》2022年第4期364-375,共12页Chinese Science Bulletin

基　　金：宁夏回族自治区自然科学基金(2019AAC03147);宁夏回族自治区重点研发计划(2019BFG02026,2020BBF02006)资助。

摘　　要：有报道指出,花色苷对疾病具有一定的化学预防活性.黑果枸杞富含花色苷,具有多种生物活性.本实验前期研究发现,黑果枸杞花色苷Pt3G可通过ROS/PTEN/PI3K/Akt途径抑制前列腺癌DU145细胞的增殖.本研究针对另外2种前列腺癌细胞LNCaP和PC-3进一步探讨黑果枸杞花色苷Pt3G抑制前列腺癌细胞的作用及其相关机制.采用甲基噻唑基四唑比色法检测细胞增殖、流式细胞术和TdT介导的dUTP缺口末端标记技术(TdT-mediated dUTP nick-end labeling,TUNEL)法检测细胞凋亡以及蛋白印迹法检测相关蛋白的表达.结果表明,Pt3G以浓度依赖性方式抑制细胞增殖,其抑制LNCaP和PC-3细胞的半数抑制浓度分别为601.97和445.79μg/mL.流式细胞术和TUNEL检测发现,Pt3G能够诱导前列腺癌LNCaP和PC-3细胞凋亡,并促进细胞周期阻滞在S期.而且,不同浓度Pt3G处理前列腺癌细胞24 h后,细胞抗凋亡因子PI3K、p-PI3K、Akt、p-Akt的表达水平显著降低,而促凋亡因子Bax、Cytochrome c、caspase 3、cleaved caspase 3的表达显著升高.此外,研究还发现,Pt3G可以激活LNCaP和PC-3细胞的PTEN蛋白并刺激细胞ROS的过量产生来诱导LNCaP和PC-3细胞凋亡.综上,本研究进一步证明了黑果枸杞花色苷Pt3G可能通过ROS/PTEN/PI3K/Akt/caspase 3途径抑制LNCaP和PC-3细胞增殖并促进细胞凋亡.因此,黑果枸杞花色苷Pt3G可能是预防或治疗前列腺癌的一种潜在抗增殖剂.Lycium ruthenicum Murray(L.ruthenicum),known as“black Goji”,has been widely distributed in northwest of China.The fruit of L.ruthenicum has been used as a traditional Chinese medicine for a long time.Many modern studies indicated that the fruit of L.ruthenicum has a variety of biological activities such as antioxidant,anti-inflammatory,anti-cancer,neuroprotective,and potential prevention of cardiovascular disease.The fruit of L.ruthenicum is black in color and rich in anthocyanins.It has been reported that petunidin 3-O-[6-O-(4-O-(trans-p-coumaroyl)-α-l-rhamnopyranosyl)-β-Dglucopyranoside]-5-O-[β-D-glucopyranoside](Pt3G)is the main constituent of anthocyanins in fully ripe fruits of L.ruthenicum.In our previous study,we found that the Pt3G from the fruit of L.ruthenicum could inhibit the proliferation of prostate cancer DU145 cells through ROS/PTEN/PI3K/Akt pathway.To further explore the mechanisms underlying the inhibitory effect of Pt3G on proliferation of prostate cancer cells,the prostate cancer LNCaP and PC-3 cells were used in this study.MTT assay was used to determine the proliferation rate of cells treated with different concentrations of Pt3G solution.To verify whether the inhibitory effect of Pt3G on the proliferation of prostate cancer cell is due to apoptosis,flow cytometric analysis and TUNEL assay were used to detect cell apoptosis,and the Western Blot analysis was used to detect the expression of apoptosis related proteins.The mitochondrial membrane potential in LNCaP and PC-3 cells was analyzed with JC-1 fluorescent dye.Our results showed that Pt3G inhibited the proliferation of LNCaP and PC-3 cells in a concentration dependent manner.The half inhibitory concentrations(IC50)of Pt3G against LNCaP and PC-3 cells were 601.97 and 445.79μg/mL at 24 h after treatment,respectively.Flow cytometry and TUNEL results showed that the mean percentages of apoptotic cells were significantly increased in the Pt3G treated LNCaP and PC-3 cells as compared to the control group.Furthermore,Pt3G could lead to

关 键 词：黑果枸杞 花色苷 细胞凋亡 ROS/PTEN/PI3K/Akt LNCaP和PC-3细胞 

分 类 号：R285[医药卫生—中药学]