The Affiliated Hospital of Guilin Medical College, Guilin 541001, P. R. ChinaAbstract:Objective To investigate the antifibrotic action and mechanism of artesunate furtherly. Methods Human embryonic lung fibroblasts (HELF) ceil lines HFL-I were cultured to log phase in vitro and divided into 4 observed groups and control group, the former 4 groups were treated with artesunate at the mass concentration of 4, 8, 16, 32 mg/L, the control group were given equivalent DMEM. The cell proliferation activity was determined by CCK-g assay, the cell cycle and apoptosis ratio were detected by flow eytometry (FCM), the expression of Survivin mRNA was detected by RT-PCR. Results Compared with the control group, percentage of G0 ? G1 stage cells and the apoptosis ratio in observed groups increased significantly, A value and the expression of Survivin mRNA decreased significantly, and remarkable differences were also seen among observed groups (P< 0.05). Conclusion Artesunate can resist pulmonary fibrosis, and probable mechanism relate with proliferation inhibition and apoptosis induction of HFL-I through downregulating expression of Survivin mRNA.
--HFL-IWANG Chang-ruing, ZHANG Xiao-fei, TAN Ning, HUANG Lan-zhen, LIN Yun, XU Qing. Effects and mechanism of artesunate on the cell proliferation and apoptosis in human embryonic lung fibroblast cell line. --Shan Dong Yi Yao. 2010; (3): 33-35.
Inhibition of Artemisunate on Invasion of Human Colon Cancer Line SW620.
Fan Y, Zhang Y-l, Yao G-t, Li Y-k. Inhibition of Artemisunate on Invasion of Human Colon Cancer Line SW620. Shi Zhen Guo Yi Guo Yao. 2008, 19(7): 1740-1741.
After colon cancer SW620 cells were treated with different doses of Artemisunate, the growth of anchorage independence of cancer cells was studied in soft agar colony formation, and invasion ability was determined by Boyden chamber, and the protein level of intercellular adhesion molecule-1 (ICAM-1) was detected by Western blot assay. Results indicate that Artemisunate could significantly inhibit both invasion ability and anchorage independence growth in a dose-dependent manner. The level of protein of ICAM-1 was down-regulated as compared with control group. Artemisunate can inhibit the invasion of colon carcinoma cell line SW620 through down-regulating ICAM-1 expression. 1
The alkaloid Berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest.
Kim JB, Yu JH, Ko E, Lee KW, et al. Phytomedicine. 2010 May;17(6):436-40. doi: 10.1016/j.phymed.2009.08.012. Epub 2009 Oct 2.
Berberine is a pure phenanthren alkaloid isolated from the roots and bark of herbal plants such as Berberis, Hydrastis canadensis and Coptis chinensis. Berberine has been established to inhibit the growth of breast cancer cells, but its effects on the drug resistance and anoikis-resistance of breast cancer cells have yet to be elucidated. Anoikis, or detachment-induced apoptosis, may prevent cancer progression and metastasis by blocking signals necessary for survival of localized cancer cells.
Resistance to anoikis is regarded as a prerequisite for metastasis; however, little is known about the role of berberine in anoikis-resistance. We established anoikis-resistant cells from the breast cancer cell lines MCF-7 and MDA-MB-231 by culturing them on a Poly-Hema substratum. We then investigated the effects of berberine on the growth of these cells. The anoikis-resistant cells had a reduced growth rate and were more invasive than their respective adherent cell lines. The effect of berberine on growth was compared to that of doxorubicine, which is a drug commonly used to treat breast cancer, in both the adherent and anoikis-resistant cell lines. Berberine promoted the growth inhibition of anoikis-resistant cells to a greater extent than doxorubicine treatment.
Treatment with berberine-induced cell cycle arrest at G0/G1 in the anoikis-resistant MCF-7 and MDA-MB-231 cells as compared to untreated control cells. In summary, these results revealed that berberine can efficiently inhibit growth by inducing cell cycle arrest in anoikis-resistant MCF-7 and MDA-MB-231 cells.
Further analysis of these phenotypes is essential for understanding the effect of berberine on anoikis-resistant breast cancer cells, which would be relevant for the therapeutic targeting of breast cancer metastasis.
Safety and efficacy field study of artesunate for dogs with non-resectable tumours
Rutteman GR, Erich SA, Mol JA, Spee B, Grinwis GC, Fleckenstein L, London CA, Efferth T. Anticancer Res. 2013 May;33(5):1819-27.
The anti-malarial drug artesunate has shown anticancer activity in vitro and in preliminary animal experiments, but experience in patients with cancer is very limited. Pre-clinical studies in dogs indicated morbidity at high dosage levels. This study evaluated the effects of artesunate in canine cancer cell lines and in canine cancer patients. Four canine cell lines were tested in vitro for sensitivity towards artesunate and dihydroartemisinin (DHA; active metabolite of artesunate). The half-maximal inhibitory concentration (IC50) values for artesunate or DHA were 2-60 μM in three cell lines, while one cell line was much less sensitive to artesunate (IC50 337 μM) than to DHA (IC50 50 μM). A safety/efficacy field study with artesunate was conducted in 23 dogs with non-resectable tumours. Artesunate was administered for 7-385 days at a dosage of 651-1178 (median 922) mg/m(2). No neurological or cardiac toxicity was observed and seven dogs exhibited no adverse effects at all. Fever and haematological/gastrointestinal toxicity, mostly transient, occurred in 16 dogs. One dog died from pneumonia. Plasma artesunate and DHA levels fell below the limit of detection within 8-12 h after artesunate administration, while levels after two hours were close to 1 μM. Artesunate produced a long-lasting complete remission in one case of cancer and short-term stabilization of another seven cases.
Artesunate induces apoptosis via a Bak-mediated caspase-independent intrinsic pathway in human lung adenocarcinoma cells.
Zhou C, Pan W, Wang XP, Chen TS. J Cell Physiol. 2012 Dec;227(12):3778-86. doi: 10.1002/jcp.24086.
This report is designed to explore the exact molecular mechanism by which artesunate (ART), a semisynthetic derivative of the herbal antimalaria drug artemisinin, induces apoptosis in human lung adenocarcinoma (ASTC-a-1 and A549) cell lines. ART treatment induced ROS-mediated apoptosis in a concentration- and time-dependent fashion accompanying the loss of mitochondrial potential and subsequent release of Smac and AIF indicative of intrinsic apoptosis pathway. Blockage of casapse-8 and -9 did not show any inhibitory effect on the ART-induced apoptosis, but which was remarkably prevented by silencing AIF. Of the utmost importance, ART treatment induced the activation of Bak but not Bax, and silencing Bak but not Bax remarkably inhibited ART-induced apoptosis and AIF release. Furthermore, although ART treatment did not induced a significant down-regulation of voltage-dependent anion channel 2 (VDAC2) expression and up-regulation of Bim expression, silencing VDAC2 potently enhanced the ART-induced Bak activation and apoptosis which were significantly prevented by silencing Bim. Collectively, our data firstly demonstrate that ART induces Bak-mediated caspase-independent intrinsic apoptosis in which Bim and VDAC2 as well as AIF play important roles in both ASTC-a-1 and A549 cell lines, indicating a potential therapeutic effect of ART for lung cancer.
Artesunate induces oxidative DNA damage, sustained DNA double-strand breaks, and the ATM/ATR damage response in cancer cells.
Berdelle N, Nikolova T, Quiros S, Efferth T, Kaina B. Mol Cancer Ther. 2011 Dec;10(12):2224-33. doi: 10.1158/1535-7163.MCT-11-0534. Epub 2011 Oct 13.
Artesunate, the active agent from Artemisia annua L. used in the traditional Chinese medicine, is being applied as a first-line drug for malaria treatment, and trials are ongoing that include this drug in cancer therapy. Despite increasing interest in its therapeutic application, the mode of cell killing provoked by artesunate in human cells is unknown. Here, we show that artesunate is a powerful inducer of oxidative DNA damage, giving rise to formamidopyrimidine DNA glycosylase-sensitive sites and the formation of 8-oxoguanine and 1,N6-ethenoadenine. Oxidative DNA damage was induced in LN-229 human glioblastoma cells dose dependently and was paralleled by cell death executed by apoptosis and necrosis, which could be attenuated by radical scavengers such as N-acetyl cysteine. Oxidative DNA damage resulted in DNA double-strand breaks (DSB) as determined by γH2AX foci that colocalized with 53BP1. Upon chronic treatment with artesunate, the level of DSB continuously increased over the treatment period up to a steady-state level, which is in contrast to ionizing radiation that induced a burst of DSB followed by a decline due to their repair. Knockdown of Rad51 by short interfering RNA and inactivation of DNA-PK strongly sensitized glioma cells to artesunate. These data indicate that both homologous recombination and nonhomologous end joining are involved in the repair of artesunate-induced DSB. Artesunate provoked a DNA damage response (DDR) with phosphorylation of ATM, ATR, Chk1, and Chk2. Overall, these data revealed that artesunate induces oxidative DNA lesions and DSB that continuously increase during the treatment period and accumulate until they trigger DDR and finally tumor cell death.
The Expression Of TGF-β1,SMAD3,CDC25A,CDC25B And In Esophageal Cancer And The Effects Of Artesunate Against Esophageal Cancer
Objective: To detect the express of CDC25A,CDC25B,SMAD3,TGF-β1 in esophageal cancer, approaching the effects of Artesunate against esophageal cancer and related mechanism, searching the new medicine for the therapy of esophagealcancer.Methods1 Researching esophageal cancer,adjacent mucosa to esophageal carcinoma and normal esophageal mucosa at surgical margin of esophageal carcinoma1.1 To study the mRNA of CDC25A,CDC25B,SMAD3,TGF-β1 in 44 example esophageal carcinoma, paired adjacent mucosa to esophageal carcinoma and normal esophageal mucosa at surgical margin of esophageal carcinoma tissues by RT-PCR. The 44 example was all diagnosed by pathology.1.2 To study 58 example primarily esophageal carcinoma and normal esophageal mucosa,to research the cell cycle,apoptosis and the protein of CDC25A,CDC25B,SMAD3.All examples were diagnosed by pathology.2 To study the role of CDC25A,CDC25B,SMAD3,TGF-β1 in the course of Artesunate against esophageal carcinoma. To set up esophageal carcinoma nude mouse animal model, thity in all,divided them for five groups randomly.Every nude mouse injected 6×106.200ul-1 Eca109 cells in left preepipodite, growing tumor in a week ,the mean size of tumor was 0.4mm3, starting to inject drug ,tow weeks ,one course of treatment was one week ,the first group injected Artesunate 100mg.kg-1,the second group injected Artesunate 200mg.kg-1,the third group injected Artesunate 300mg.kg-1,the forth group injected Cisplatin 3mg.kg-1as masculine control group ,the fifth group inject- ted Sodium Chloride as negative control group.injected drugs in abdominal cavity.tow weeks later,killed all of the nude mouse. Taking out of the tumor,divided two share,one share tested cellcycle,Apoptosis and the protein of CDC25A,SMAD3 using Flow cytometry.
The other tested the CDC25A,CDC25B,SMAD3 and TGF-β1of the tumor by PT-PCR.GAPDH as internal reference.3 To make use of spss11.5 software carrying out statistical treatment.Using x±s expressed Test data,To compare different groups using ANOVA,To compare two groups by T test.α=0.05 as the size of test.when P<0.05,having statistical significance.Results1 The express of CDC25A,CDC25B,SMAD3 and TGF-β1 mRNA in different pathological changes of esophageal tissues1.1 The express of CDC25A mRNA in different pathological changes of esophageal tissuesThe express of CDC25A mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.426±0.210,0.494±0.205,0.705±0.185,The express of CDC25A mRNA was higher in esophageal carcinoma than others(P<0.05).1.2 The express of CDC25B mRNA in different pathological changes of esophageal tissuesThe express of CDC25B mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.3832±01440,0.4550±0.1744,0.5802±0.4188,The express of CDC25B mRNA was higher in esophageal carcinoma than others(P<0.05).1.3 The express of SMAD3 mRNA in different pathological changes of esophageal tissuesThe express of SMAD3 mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.623±0.281,0.613±0.278,0.479±0.251,The express of SMAD3 mRNA was lower in esophageal carcinoma than others(P<0.05).1.4 The express of TGF-β1 mRNA in different pathological changes of esophageal tissuesThe express of TGF-β1 mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.5963±0.2437,0.5592±0.2575,0.4555±0.1585,The express of TGF-β1 mRNA was lower in esophageal carcinoma than others(P<0.05).2 To study the cell cycle and apoptosis in different pathological changes of esophageal tissues by FCMThe apoptosis of esophageal carcinoma was lower than normal esophageal mucosa(P<0.05). The apoptosis of esophageal carcinoma was (8.10±3.57)%, The apoptosis of normal esophageal mucosa was (9.53±2.26)%.
The G1 stage of normal esophageal mucosa was higher than esophageal carcinoma(P<0.05). The G1 stage of normal esophageal mucosa was(86.27±3.72)%,The G1 stage of esophageal carcinoma was(75.07±11.01)%.The proliferation index of normal esophageal mucosa was lower than esophageal carcinoma(P<0.05). The proliferation index of normal esophageal mucosa was(14.49±3.83)%, The proliferation index of esophageal carcinoma was(24.92±11.01)%.3 To study the protein of CDC25A,CDC25B and SMAD3 in different pathological changes of esophageal tissues by FCMThe protein of CDC25A and CDC25B in esophageal carcinoma was higher than normal esophageal mucosa (P<0.05), The protein of SMAD3 in esophageal carcinoma was lower than normal esophageal mucosa(P<0.05).4 The inhibition effects of Artesunate(Art) on the growth of Eca109 transplated subcutaneous tumors in nude miceThe volume and weight of Eca109 transplated subcutaneous tumors in every experiment group was lower than negative control group(P<0.05). The volume of Eca109 transplated subcutaneous tumors in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was(0.66±0.29)cm3,(0.28±0.22)cm3,(0.61±0.38)cm3 respectively. The weight of Eca109 transplated subcutaneous tumors in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was (0.66±0.27)g,(0.37±0.16)g,(0.65±0.19)g respectively. The volume of Eca109 transplated subcutaneous tumors in DDP group was(0.21±0.07)cm3, The weight of Eca109 transplated subcutaneous tumors in DDP group was (0.33±0.05)g, The volume of Eca109 transplated subcutaneous tumors in normal sodium group was(1.00±0.30)cm3, The weight of Eca109 transplated subcutaneous tumors in normal sodium group was(0.94±0.31)g. The inhibition rate of volume in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was 34%,72%,39% respectively, The inhibition rate of weight in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was 29.79%,60.64%,30.85% respectively. The inhibition rate of volume in DDP group was 79%,The inhibition rate of weight in DDP group was 64.89%.5 The express of CDC25A,CDC25B,SMAD3,TGF-β1 mRNA in Eca109 transplated subcutaneous tumors in nude mice5.1 The express of CDC25A mRNA100mg.kg-1Art group was 0.8383±0.0454,200mg.kg-1Art group was 0.7300±0.0583,300mg.kg-1Art group was 0.8517±0.0483,DDP group was 0.6850±0.1436,normal sodium group was 0.9650±0.0635. normal sodium group was higher than others(P<0.05). 5.2 The express of CDC25BmRNA100mg.kg-1Art group was 0.5036±0.0041, 200mg.kg-1Art group was 0.4556±0.0353,300mg.kg-1Art group was 0.5024±0.0047,DDP group was 0.4406±0.0354,normal sodium group was 0.5389±0.0407. normal sodium group was higher than others(P<0.05).5.3 The express of SMAD3 mRNA100mg.kg-1Art group was 0.7865±0.0421, 200mg.kg-1Art group was 0.8909±0.0832,300mg.kg-1Art group was 0.7915±0.0582,DDP group was 0.8914±0.0410,normal sodium group was 0.6862±0.0518. normal sodium group was lower than others(P<0.05).5.4 The express of TGF-β1 mRNA100mg.kg-1Art group was 0.6703±0.0193, 200mg.kg-1Art group was 0.7439±0.0546,300mg.kg-1Art group was 0.6777±0.0284,DDP group was 0.7083±0.0628,normal sodium group was 0.6091±0.0405. normal sodium group was lower than others(P<0.05).6 The express of CDC25A,SMAD3 protein in Eca109 transplated subcutaneous tumors in nude miceCDC25A protein: 100mg.kg-1 Art group was 513.5761±18.7518, 200mg.kg-1 Art group was 470.0678±17.2265, 300mg. kg-1Art group was 512.5522±13.3181,DDP group was 484.5414±25.9349,normal sodium group was 552.1977±22.0075. normal sodium group was higher than others(P<0.05). SMAD3 protein: 100mg.kg-1Art group was 418.2193±11.3027, 200mg.kg-1Art group was 454.6632±33.9175, 300mg. kg-1Art group was 418.9937±31.1770,DDP group was 456.5815±37.2966,normal sodium group was 371.4166±23.2261. normal sodium group was lower than others (P<0.05).7
The cell cycle of Eca109 transplated subcutaneous tumors in nude miceThe cell cycle was stopped in G0- G1 stage(66.3%)in Art groups(P<0.05). The S stage in normal sodium group was 42.9%,higher than others,to show that tumor cells grew vigorously.Conclusion1. CDC25A mRNA and protein in esophageal carcinoma was higher than normal esophageal mucosa. CDC25B mRNA and protein in esophageal carcinoma was higher than normal esophageal mucosa.SMAD3 mRNA and protein in esophageal carcinoma was lower than normal esophageal mucosa. TGF-β1 mRNA in esophageal carcinoma was lower than normal esophageal mucosa.these gene and protein maybe the molecule mechanism to cause esophageal carcinoma.2. The apoptosis rate in normal esophageal mucosa was higher than esophageal carcinoma, G1 stage in normal esophageal mucosa was higher than esophageal carcinoma, s stage in esophageal carcinoma was higher than normal esophageal mucosa ,to show that tumor cells grow vigorously.3. Artesunate can stop the growth of Eca109 transplated subcutaneous tumors in nude mice,the middle density Artesunate had the strongest effect in stopping the growth of Eca109 transplated subcutaneous tumors in nude mice in three Artesunate groups.4. Artesunate can stop the transplated subcutaneous tumors cell in G1 stage,so the role of Artesunate against esophageal carcinoma maybe relate to cell cycle blockage.5. Artesunate can increase the expression of SMAD3 and TGF-β1,reduce the expression of CDC25A and CDC25B.so the role of Artesunate against esophageal carcinoma maybe relate to CDC25A,CDC25B,SMAD3 and TGF-β1.
Keywords/Search Tags: esophageal squamous cell cancer, CDC25A, SMAD3, Athymic mouse
1 Expression Of CDC25A, CDC25B, TGF-beta, Smad3 In Esophageal Squamous Cell Carcinoma And Their Role In Antitumor Effect Of Artesunate
2 Expression Of CDC25A And TGF-β/Smad3 Signal Transducer In Esophageal Carcinoma And Their Role In Antitumor Activity Of Artesunate
3 The Role Of S100A4 In The Infiltration, Metastasis And Its Influence To The Growth Of Esophageal Squamous Cell Carcinoma
4 Comparative Proteomic Analysis Of Esophageal Squamous Cell Carcinoma From High-incidence Area Of Esophageal Cancer
5 Inhibitory Effect Of SiRNA Targeting HIF-1α On Esophageal Squamous Cancer Cells Eca-109 In Vitro And In Vivo
6 1. Gut-enriched Krüppel Like Factor (GKLF) Is Associated With Cell Differentiation And Regulates Keratin 13 In Human Esophageal Squamous Cell Carcinoma 2. The Role Of P53 And MDM2 Polymorphisms In The Risk Of Esophageal Squamous Cell Carcinoma
7 Constructing Of Cloned Esophageal Squamous Cancer Cell Strain With Hypoxia-Inducible Factor 1α Gene Silencing And Initial Study On The Role Of HIF-1α Gene In Esophageal Squamous Cancer Cell
8 The Effect Of Smad7sirna On Generation And Invasion In Escc Cell Line Ec-1
9 Expression And Preliminary Function Analysis Of AMF Gene In Esophageal Squamous Cell Cancer Cells
10 Effect Of Calcitriol Combinating Ddp On The HEC-1-A Transplantation Tumor Of Athymic Mouse