Objective To study the antitumor activity of Xerophilusin G on S180 cells,and Its mechanism. Methods Modified MTT assay was used to test the effect of Xerophilusin G on the proliferation of S180 tumor cell strain. The influences on tumor growth and immune organs of mice with transplanted sarcoma (S180) were observed. The cell cycle of S180 cell lines and mouse sarcoma (S180) was analyzed by flow cytometry. The lymphocyte proliferation activity of spleen stimulating was tested. The level of IL-2 in serum of mice with transplanted sarcoma (S180) was measured by ELISA. Results The IC50 of Xerophilusin G in S180 cell lines was 19.80 μg·mL-1, the LD50 in mouse for Xerophilusin G was 121.11 mg·kg-1 through intraperitoneal injection. The tumor inhibition rate of Xerophilusin G was 32.11% and 41.60%, respectively at the doses of 3 and 6 mg·kg-1 (P<0.05). Compared with the control, the thymus, kidney and cardiac index were decreased. The cell proportion at G0/G1 phase of mouse sarcoma (S180) was increased. T and B cell proliferation activities in tumor-bearing mice were enhanced (P<0.05). As compared with control group, the serum level of IL-2 was decreased 90.9% and 77.1% in low- and medium-dose groups, respectively (P<0.05). ConclusionXerophilusin G has remarkable effects in sarcoma (S180) bearing mice. The antitumor mechanism of Xerophilusin G might be related with G0/G1 phase arrest of mouse sarcoma (S180) cells and enhancing the activity of T and B cell but not related with increasing the secreting of IL-2.
旱生香茶菜是云南特有的香茶菜物种,药用其枝叶,其味苦,性寒,具有清热解毒、活血化瘀、抗菌消炎以及抗肿瘤等功效[1]。该植物中富含有对映-贝壳杉烷型二萜化合物,其中旱生香茶菜素G(Xerophilusin G)含量最多,约占总二萜的2.5%。本课题组前期研究旱生香茶菜总二萜(the total diterpenoids of Isodon Xerophilus,IXD)对人肿瘤细胞株K562、CA、CNE、BGC-823、MKN-28、A549、T-24等体外细胞毒活性,发现它能明显抑制肿瘤细胞增殖,且抗瘤谱较广[2];IXD 对人肝癌细胞株CA体外有较强的细胞毒活性,对荷CA 裸鼠移植瘤生长具有一定的抑制作用[3]。为从旱生香茶菜总二萜中发现新的抗肿瘤先导化合物,笔者以荷S180肉瘤小鼠为模型拟通过对旱生香茶菜素G抗肿瘤活性及其机制进行研究。
1 材料与方法
1.1 瘤株与实验动物
小鼠肉瘤细胞株S180,引自中国科学院上海药物研究所;美国癌症研究所(Institute of Cancer Research,ICR)小鼠,无特定病原体(SPF)级,雌雄各半,体质量18~22 g,由昆明医学院实验动物中心提供,实验动物生产许可证号:SCXK(滇)2005-0008。
表4
6组荷S180肉瘤小鼠脾细胞增殖及血清中IL-2含量的比较
Tab.4
Comparison of splenocyte proliferation and the serum level of IL-2 among six groups of mice bearing S180 tumor ヌ±s,n=4
组别
剂量/ (mg·kg-1)
T淋巴细胞增殖率 %
B淋巴细胞增殖率
IL-2/ (pg·mL-1)
空白对照组
—
8.81±4.17
7.76±1.64
11.78±0.12
阴性对照组
—
-17.75±3.85
4.25±2.87
10.31±0.76
环磷酰胺组
30
2.19±1.04
-8.66±3.80
3.07±0.10*1
旱生香茶菜素G
小剂量
3
24.51±8.94*2
35.43±9.42*2
0.94±0.10*2
中剂量
6
34.27±10.25*2
50.66±15.31*2
2.36±0.14*2
大剂量
12
32.56±14.73*2
130.36±16.27*2
8.12±0.16
F
1.870
2.350
4.263
P
<0.05
<0.05
<0.05
Compared with blank control group,*1P<0.05;compared with negative control group,*2P<0.05
与空白对照组比较,*1P<0.05;与阴性对照组比较,*2P<0.05
表4
6组荷S180肉瘤小鼠脾细胞增殖及血清中IL-2含量的比较
Tab.4
Comparison of splenocyte proliferation and the serum level of IL-2 among six groups of mice bearing S180 tumor ヌ±s,n=4
SUN HD,HUANG SX,HAN QB.Diterpenoids from Isodon species and their biological activities[J].,2006,23(5),673-698.
ABSTRACT Isodon species (Labiatae) are widely distributed plants, many of which are used in folk medicine. Over the past twenty years, they have received considerable phytochemical and biological attention. Thestructures of their many diterpenoids constituents, especially those with an ent-kaurane skeleton, have been elucidated. The significant phytochemical and pharmacological diterpenoids form the subject of this review. There are 290 references.
目的:研究旱生香茶菜总二萜(Diterpenoids of Isodon Xerophilus,IXD)体外对10株人源性肿瘤细胞增殖的影响。方法:采用改良MTf法或SRB法测试其在体外对10株人肿瘤细胞增殖的影响。结果:IXD对K562、CA、CNE、BGC、MKN、A549、T-24七株人肿瘤细胞增殖的IC加值分别为5.42、6.17、11.05、19.20、24.00、24.04、25.48μg/mL。均小于30.00μg/mL;对SKOV3、Helm、GLC三株人肿瘤细胞增殖的IC50值分别为735.3、740.5、2.13×10^8μg/mL。均大于30.00μg/mL。结论:IXD在体外显示较强且广谱的细胞毒活性。
Four 7,20-epoxy ent -kaurane diterpenoids, xerophilusins G ( 1 ) and I–K ( 2 – 4 ), were isolated from the leaves of Isodon xerophilus , along with four known ones, enanderianin C ( 5 ), rosthorin A ( 6 ), longikaurin B ( 7 ), and rabdoternin D ( 8 ). Their structures were determined primarily using NMR spectroscopic techniques. The structure and stereochemistry of 3 were confirmed by X-ray crystallography. Compounds 4 and 7 exhibited broad cytotoxicity against four kinds of human tumor cells (K562, HL-60, HCT, and MKN-28 cells) in the range of 2.23–15.35 and 0.30–8.61 μg/ml, respectively.
DIAZ MS,TARRADO CM,MIRANDAA,et al.Targeting cell cycle regulation in cancer therapy[J].,2013,138(2):255-271.
Abstract Cell proliferation is an essential mechanism for growth, development and regeneration of eukaryotic organisms; however, it is also the cause of one of the most devastating diseases of our era: cancer. Given the relevance of the processes in which cell proliferation is involved, its regulation is of paramount importance for multicellular organisms. Cell division is orchestrated by a complex network of interactions between proteins, metabolism and microenvironment including several signaling pathways and mechanisms of control aiming to enable cell proliferation only in response to specific stimuli and under adequate conditions. Three main players have been identified in the coordinated variation of the many molecules that play a role in cell cycle: i) The cell cycle protein machinery including cyclin-dependent kinases (CDK)-cyclin complexes and related kinases, ii) The metabolic enzymes and related metabolites and iii) The reactive-oxygen species (ROS) and cellular redox status. The role of these key players and the interaction between oscillatory and non-oscillatory species has proved essential for driving the cell cycle. Moreover, cancer development has been associated to defects in all of them. Here, we provide an overview on the role of CDK-cyclin complexes, metabolic adaptations and oxidative stress in regulating progression trough each cell cycle phase and transitions between them. Thus, new approaches for the design of innovative cancer therapies targeting crosstalk between cell cycle simultaneous events are proposed.
ZHANG XH,LIU YX,JIAM,et al.Oridonin inhibits tumor growth in glioma by inducing cell cycle arrest and apoptosis[J].,2014,60(6):29-36.
ABSTRACT Glioma is the most common malignant intracranial tumors. Despite newly developed therapies, these treatments mainly target oncogenic signals, and unfortunately, fail to provide enough survival benefit in both human patients and mouse xenograft models, especially the first—generation therapies. Oridonin is purified from the Chinese herb Rabdosia rubescens and considered to exert extensive anti—cancer effects on human tumorigenesis. In this study, we systemically investigated the role of Oridonin in tumor growth and the underlying mechanisms in human glioma. We found that Oridonin inhibited cell proliferations in a dose— and time—dependent manner in both glioma U87 and U251 cells. Moreover, these anti—cancer effects were also confirmed in a mouse model bearing glioma. Furthermore, cell cycle arrest in S phase was observed in Oridonin—mediated growth inhibition by flow cytometry. Cell cycle arrest in S phase led to eventual cell apoptosis, as revealed by Hoechst 33342 staining and annexin V/PI double—staining. The cell apoptosis might be accomplished through a mitochondrial manner. In all, we were the first to our knowledge to report that Oridonin could exert anti—cancer effects on tumor growth in human glioma by inducing cell cycle arrest and eventual cell apoptosis. The identification of Oridonin as a critical mediator of glioma growth may potentiate Oridonin as a novel therapeutic strategies in glioma treatments.
GAO SY,LIJ,QU XY,et al.Downregulation of Cdk1 and cyclinB1 expression contributes to oridonin-induced cell cycle arrest at G2/M phase and growth inhibition in SGC-7901 gastric cancer cells[J].,2014,15(15):6437-6441.
Abstract Background: Oridonin isolated from Rabdosia rubescens, a plant used to treat cancer in Chinese folk medicine, is one of the most important antitumor active ingredients. Previous studies have shown that oridonin has anti- tumor activities in vivo and in vitro, but little is known about cell cycle effects of oridonin in gastric cancer. Materials and methods: MTT assay was adopted to detect the proliferation inhibition of SGC-7901 cells, the cell cycle was assessed by flow cytometry and protein expression by Western blotting. Results: Oridonin could inhibit SGC-7901 cell proliferation, the IC50 being 15.6 M, and blocked SGC-7901 cell cycling in the G2/M phase. The agent also decreased the protein expression of cyclinB1 and CDK1. Conclusions: Oridonin may inhibit SGC-7901 growth and block the cells in the G2/M phase by decreasing Cdk1 and cyclinB1 proteins.
LIL,YUE GG,LAU CB,et al.Eriocalyxin B induces apoptosis and cell cycle arrest in pancreatic adenocarcinoma cells through caspase and p53 dependent pathways[J].,2012,262(1):80-90.
Abstract Pancreatic cancer is difficult to detect early and responds poorly to chemotherapy. A breakthrough in the development of new therapeutic agents is urgently needed. Eriocalyxin B (EriB), isolated from the Isodon eriocalyx plant, is an ent-kaurane diterpenoid with promise as a broad-spectrum anti-cancer agent. The anti-leukemic activity of EriB, including the underlying mechanisms involved, has been particularly well documented. In this study, we demonstrated for the first time EriB's potent cytotoxicity against four pancreatic adenocarcinoma cell lines, namely PANC-1, SW1990, CAPAN-1, and CAPAN-2. The effects were comparable to that of the chemotherapeutic camptothecin (CAM), but with much lower toxicity against normal human liver WRL68 cells. EriB's cytoxicity against CAPAN-2 cells was found to involve caspase-dependent apoptosis and cell cycle arrest at the G2/M phase. Moreover, the p53 pathway was found to be activated by EriB in these cells. Furthermore, in vivo studies showed that EriB inhibited the growth of human pancreatic tumor xenografts in BALB/c nude mice without significant secondary adverse effects. These results suggest that EriB should be considered a candidate for pancreatic cancer treatment.
MONJA AM,TIETZE JK,GROSS SK,et al.Bystander activation and anti-tumor effects of CD8+ T cells following Interleukin-2 based immunotherapy is independent of CD4+ T cell help [J].,2014,9(8):e102709.
ABSTRACT We have previously demonstrated that immunotherapy combining agonistic anti-CD40 and IL-2 (IT) results in synergistic anti-tumor effects. IT induces expansion of highly cytolytic, antigen-independent "bystander-activated" (CD8+CD44high) T cells displaying a CD25-NKG2D+ phenotype in a cytokine dependent manner, which were responsible for the anti-tumor effects. While much attention has focused on CD4+ T cell help for antigen-specific CD8+ T cell expansion, little is known regarding the role of CD4+ T cells in antigen-nonspecific bystander-memory CD8+ T cell expansion. Utilizing CD4 deficient mouse models, we observed a significant expansion of bystander-memory T cells following IT which was similar to the non-CD4 depleted mice. Expanded bystander-memory CD8+ T cells upregulated PD-1 in the absence of CD4+ T cells which has been published as a hallmark of exhaustion and dysfunction in helpless CD8+ T cells. Interestingly, compared to CD8+ T cells from CD4 replete hosts, these bystander expanded cells displayed comparable (or enhanced) cytokine production, lytic ability, and in vivo anti-tumor effects suggesting no functional impairment or exhaustion and were enriched in an effector phenotype. There was no acceleration of the post-IT contraction phase of the bystander memory CD8+ response in CD4-depleted mice. The response was independent of IL-21 signaling. These results suggest that, in contrast to antigen-specific CD8+ T cell expansion, CD4+ T cell help is not necessary for expansion and activation of antigen-nonspecific bystander-memory CD8+ T cells following IT, but may play a role in regulating conversion of these cells from a central memory to effector phenotype. Additionally, the expression of PD-1 in this model appears to be a marker of effector function and not exhaustion.
Downregulation of Cdk1 and cyclinB1 expression contributes to oridonin-induced cell cycle arrest at G/M phase and growth inhibition in SGC-7901 gastric cancer cells