Singh Shashank K, Shanmugavel M, Kampasi Himani, Singh Reena, Mondhe D M, Rao J Madusudana, Adwankar M K, Saxena A K, Qazi G N
Pharmacology Division, Regional Research Laboratory (Council of Scientific and Industrial Research), Jammu Tawi, India.
Planta Med. 2007 Jun;73(6):519-26. doi: 10.1055/s-2007-967185. Epub 2007 May 30.
An isolate "CD lignan mixture" comprising lignans from stem wood of Cedrus deodara consisted of (-)-wikstromal (75 - 79%), (-)-matairesinol (9 - 13%) and benzylbutyrolactol (7 - 11%) and was studied for its in vitro cytotoxicity against human cancer cell lines. The in vivo anticancer activity of CD lignan mixture was studied using Ehrlich ascites carcinoma and colon carcinoma (CA-51) models in mice. Its effect was also studied on annexin V binding, intracellular caspases and DNA fragmentation to gain insight into the mode of action. In vitro cytotoxicity studies showed significant dose-dependent effects against several cancer cell lines from different tissues such as breast, cervix, neuroblastoma, colon, liver, and prostate at 10, 30 and 100 microg/mL. The IC (50) values varied from 16.4 ng/mL to 116.03 microg/mL depending on the cell line. Comparative data of IC (50) values of CD lignan mixture showed a synergistic effect in comparison to the individual molecules, i. e., (-)-matairesinol, (-)-wikstromol present in CD lignan mixture . CD lignan mixture had the most pronounced effect on CNS cell lines followed by colon. The tumor regression observed with Ehrlich ascites carcinoma and CA-51 was 53% and approximately 54%, respectively, when CD lignan mixture was given at 300 mg/kg, I. P. for nine days in the Ehrlich ascites carcinoma model and 400 mg/kg, I. P. for the same period in the CA-51 model. It was comparable with 5-fluorouracil at 22 mg/kg and 20 mg/kg, respectively. CD lignan mixture at 10, 30 and 100 microg/mL increased the percentage of annexin V positive HL-60 cells to 1.9 - 17.18% as compared to control (1.04%). In K562 cells CD lignan mixture at 10, 30 or 100 microg/mL and staurosporine (1 microM) showed 9.13%, 11.38%, 17.22% and 28.07% intracellular caspases activation, respectively. A distinct DNA laddering pattern was observed for treatment with the CD lignan mixture in HL-60, K562 (30 microg/mL and 100 microg/mL) and MOLT-4 cells (30 microg/mL) after 24 h incubation. DNA cell cycle analysis indicated that CD lignan mixture at 10, 30 and 100 microg/mL increased the content of hypodiploid (sub G(1) phase) cells when compared to control (2.55, 5.4 and 6.25% vs. 0.27%). The present study indicates that CD lignan mixture has cytotoxic potential against human cancer cell lines. It has the ability to induce tumor regression in vivo. It induces apoptosis as indicated by annexin V positive cells, induction of intracellular caspases, DNA fragmentation and DNA cell cycle analysis.
一种从喜马拉雅雪松茎木中提取的木脂素混合物“CD木脂素混合物”，由（-）-维克斯特罗醇（75 - 79%）、（-）-罗汉松脂素（9 - 13%）和苄基丁内酯（7 - 11%）组成，并对其针对人类癌细胞系的体外细胞毒性进行了研究。利用艾氏腹水癌和结肠癌（CA - 51）小鼠模型研究了CD木脂素混合物的体内抗癌活性。还研究了其对膜联蛋白V结合、细胞内半胱天冬酶和DNA片段化的影响，以深入了解其作用方式。体外细胞毒性研究表明，在10、30和100微克/毫升浓度下，对来自不同组织（如乳腺、宫颈、神经母细胞瘤、结肠、肝脏和前列腺）的几种癌细胞系具有显著的剂量依赖性效应。IC（50）值在16.4纳克/毫升至116.03微克/毫升之间，具体取决于细胞系。CD木脂素混合物IC（50）值的比较数据显示，与单个分子（即CD木脂素混合物中存在的（-）-罗汉松脂素、（-）-维克斯特罗醇）相比具有协同效应。CD木脂素混合物对中枢神经系统细胞系的影响最为明显，其次是结肠细胞系。在艾氏腹水癌模型中，当以300毫克/千克腹腔注射给药9天时，CD木脂素混合物对艾氏腹水癌的肿瘤消退率为53%；在CA - 51模型中，以400毫克/千克腹腔注射给药相同时间时，肿瘤消退率约为54%。这分别与22毫克/千克和20毫克/千克的5 - 氟尿嘧啶相当。与对照（1.04%）相比，10、30和100微克/毫升的CD木脂素混合物使膜联蛋白V阳性HL - 60细胞的百分比增加到1.9 - 17.18%。在K562细胞中，10、30或100微克/毫升的CD木脂素混合物以及星形孢菌素（1微摩尔）分别显示细胞内半胱天冬酶激活率为9.13%、11.38%、17.22%和28.07%。在孵育24小时后，用CD木脂素混合物处理HL - 60、K562（30微克/毫升和100微克/毫升）和MOLT - 4细胞（30微克/毫升）后观察到明显的DNA梯状条带模式。DNA细胞周期分析表明，与对照（2.55%、5.4%和6.25%对0.27%）相比，10、30和100微克/毫升的CD木脂素混合物增加了亚二倍体（亚G（1）期）细胞的含量。本研究表明，CD木脂素混合物对人类癌细胞系具有细胞毒性潜力。它具有在体内诱导肿瘤消退的能力。如膜联蛋白V阳性细胞、细胞内半胱天冬酶的诱导、DNA片段化和DNA细胞周期分析所示，它可诱导细胞凋亡。
