Curcumin inhibits cervical cancer cell proliferation,migration and invasion by suppressing the PI3K/AKT signaling pathway via the miR-29b/KDM2A axis

In this study, we aimed to examine whether curcumin exerted its anti-tumor effects by regulating miR-29b/KDM2A in cervical cancer cells. The cell viability, migration and invasion were estimated in HeLa cervical cancer cells treated with curcumin. The effects of microRNA-29b (miR-29b) on biological behaviors of HeLa SiHa cells were also assessed. Potential target genes of miR-29b were predicted and confirmed using a luciferase reporter assay, and the effects of curcumin and miR-29b on the PI3K/AKTsignaling pathway were analyzed. Curcumin treatment inhibited cell proliferation, migration and invasion of HeLa cells (P<0.05). The miR-29b expression was promoted by curcumin treatment in HeLa cells (P<0.01), and miR-29b depletion could restore the effects of curcumin on cell proliferation, migration and invasion of HeLa cells (P<0.05). KDM2A was proved as a direct target gene of miR-29b, and the activity of the PI3K/AKT signaling could be regulated by curcumin and miR-29b (P<0.05). All the data revealed that curcumin played a protective role in cervical cancer. The proliferation, migration and invasion of cervical cancer cells were inhibited by curcumin through the miR-29b/KDM2A/PI3K/AKT pathway.     

姜黄素对人子宫内膜癌细胞增殖和凋亡的影响

目的:探讨姜黄素对人子宫内膜癌细胞(HEC-1B)增殖和凋亡的影响。方法:应用6.67~66.67μmol/L的姜黄素分别处理HEC-1B 48h后,采用MTT法检测姜黄素对HEC-1B的增殖程度的影响;流式细胞仪进行细胞周期时相分析;荧光显微镜观察细胞核形态的变化。结果:①MTT法显示培养48h后,姜黄素组的吸光度值A490nm低于对照组(P<0.01),且在一定浓度范围内呈剂量依赖性;②流式细胞仪检测显示培养48h后,姜黄素组的G2/M比例高于对照组(P<0.05),且在一定浓度范围内呈剂量依赖性;③荧光显微镜下给药组部分细胞发生凋亡形态学改变,凋亡率为10.22%~34.72%。结论:姜黄素可抑制HEC-1B的增殖,诱导细胞凋亡,其诱导凋亡的机制可能是通过将细胞阻滞在G2/M期来实现的。     

An in vitro study of liposomal curcumin: stability, toxicity and biological activity in human lymphocytes and Epstein-Barr virus-transformed human B-cells

Curcumin is a multi-functional and pharmacologically safe natural agent. Used as a food additive for centuries, it also has anti-inflammatory, anti-virus and anti-tumor properties. We previously found that it is a potent inhibitor of cyclosporin A (CsA)-resistant T-cell co-stimulation pathway. It inhibits mitogen-stimulated lymphocyte proliferation, NFkappaB activation and IL-2 signaling. In spite of its safety and efficacy, the in vivo bioavailability of curcumin is poor, and this may be a major obstacle to its utility as a therapeutic agent. Liposomes are known to be excellent carriers for drug delivery. In this in vitro study, we report the effects of different liposome formulations on curcumin stability in phosphate buffered saline (PBS), human blood, plasma and culture medium RPMI-1640+10% FBS (pH 7.4, 37 degrees C). Liposomal curcumin had higher stability than free curcumin in PBS. Liposomal and free curcumin had similar stability in human blood, plasma and RPMI-1640+10% FBS. We looked at the toxicity of non-drug-containing liposomes on (3)H-thymidine incorporation by concanavalin A (Con A)-stimulated human lymphocytes, splenocytes and Epstein-Barr virus (EBV)-transformed human B-cell lymphoblastoid cell line (LCL). We found that dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylglycerol (DMPG) were toxic to the tested cells. However, addition of cholesterol to the lipids at DMPC:DMPG:cholesterol=7:1:8 (molar ratio) almost completely eliminated the lipid toxicity to these cells. Liposomal curcumin had similar or even stronger inhibitory effects on Con A-stimulated human lymphocyte, splenocyte and LCL proliferation. We conclude that liposomal curcumin may be useful for intravenous administration to improve the bioavailability and efficacy, facilitating in vivo studies that could ultimately lead to clinical application of curcumin.     

Inhibition of curcumin on myeloid-derived suppressor cells is requisite for controlling lung cancer

Lung cancer remains the leading cause of cancer mortality. Myeloid-derived suppressor cells (MDSCs) are potent immune-suppressive cells and present in most cancer patients. Recently, several studies have shown that curcumin inhibits the expansion of MDSCs in some cancers. However, it is not clear how curcumin modulates the suppressive function of MDSCs, and whether curcumin achieves anti-tumor effects via regulating the expansion of MDSCs in lung cancer. Here, our results showed that curcumin significantly inhibited tumor growth in a Lewis lung carcinoma (LLC) isogenic tumor model. Curcumin reduced the accumulation of MDSCs in spleen and tumor tissue in LLC isogenic model. And curcumin promoted the maturation and differentiation of MDSCs in tumor tissue. Notably, curcumin inhibited the expression level of immune suppressive factors of MDSCs, arginase-1 (Arg-1) and ROS, in purified MDSCs from tumor tissue in vivo. Expectedly, curcumin also inhibited the immunosuppressive function of isolated MDSCs from tumor tissue and spleen of tumor bearing mice in vitro. Moreover, curcumin decreased the level of IL-6 in the tumor tissue and serum from LLC-bearing mice. Taken together, curcumin indeed possesses anti-cancer effect and inhibits the accumulation and function of MDSCs. And curcumin reduces the level of IL-6 in tumor-bearing mice to impair the expansion and function of MDSCs. These results suggest that inhibition of MDSCs in tumor is requisite for controlling lung cancer.     

雄黄注射液体内外抗肿瘤作用研究

目的:初步探讨雄黄注射液的体内外抗肿瘤作用。方法:采用噻唑蓝(MTT)方法观察雄黄注射液对培养的人白血病细胞株K562,人肝癌细胞株HepG-2,人胃癌细胞株MGC-803,和小鼠Lewis肺癌细胞的增殖抑制作用;建立小鼠S180腹水型肉瘤模型,采用1、2、3mg/kg剂量雄黄注射液作用于小鼠模型,观察雄黄注射液体内抗肿瘤作用。结果:MTT法显示雄黄注射液对人白血病细胞株K562,人肝癌细胞株HepG-2,人胃癌细胞株MGC-803,和小鼠Lewis肺癌细胞增殖有一定的抑制作用;灌胃给予雄黄注射液对小鼠S180腹水型肉瘤生长有一定抑制作用。结论:雄黄注射液能够抑制培养的肿瘤细胞生长,灌胃给药对S180腹水型肉瘤细胞移植的模型小鼠肿瘤增殖有明显的抑制作用。     

蟾酥脂质体注射液的抗肿瘤作用

目的探讨蟾酥脂质体注射液(CS)的抗肿瘤作用。方法体内实验采用小鼠肝癌H22、肉瘤S180荷瘤小鼠模型,考察CS的抑瘤作用;采用小鼠接种肝癌H22后,用环磷酰胺造成免疫低下模型,考察CS提高机体免疫力作用。体外实验采用MTT法,测CS对人宫颈癌HeLa细胞或人肝癌BEL-7402细胞增殖抑制率。结果CS(0.2、0.4、0.8 mg.kg-1)对2种荷瘤小鼠肿瘤增长有显著或非常显著的抑制作用(P<0.05,P<0.01);同时对机体具有明显的免疫调节作用。CS 0.01~3.00 mg.L-1对人宫颈癌HeLa细胞或肝癌BEL-7402细胞增殖有非常显著的抑制作用;CS对人宫颈癌HeLa细胞IC50为0.51 mg.L-1,95%可信限0.213 2~1.214 0 mg.L-1,CS对人肝癌BEL-7402细胞IC50为0.75 mg.L-1,95%可信限0.312 6~1.801 5 mg.L-1。结论蟾酥脂质体注射液体内、体外均有抗肿瘤活性,且具有免疫调节功能。     

薏苡仁提取物注射液对人肺腺癌的抗肿瘤作用研究

目的研究薏苡仁提取物注射液（康莱特,Kanglaite）对人肺腺癌体外及体内的抗肿瘤作用。方法运用CCK-8细胞增殖试验、划痕试验以及Transwell小室穿膜试验考察康莱特对A549细胞株的增殖、迁移及侵袭的影响;建立人肺腺癌A549裸鼠异种移植模型,考察高、中和低剂量静脉给药的康莱特单用或与顺铂合用的抗肿瘤作用。结果体外试验显示,经康莱特作用后,A549细胞的增殖、迁移和侵袭均受到明显抑制。康莱特高、中和低剂量静脉给药,对人肺腺癌A549裸鼠异种移植模型的肿瘤相对增殖率分别为34．52％、36．67％和42．47％;高、中和低剂量康莱特合并低剂量顺铂的肿瘤相对增殖率分别为26．75％、22．48％及29．82％。结论康莱特在体外及体内对人肺腺癌均有良好的抗肿瘤活性。康莱特与低剂量顺铂合用,体内抗肿瘤作用增强。     

Characterization and antitumor activity of pollen polysaccharide

The polysaccharide LBPP was extracted and isolated from the pollen of brassica napus L., and the antitumor activity was evaluated on Sarcoma 180-bearing mice and B16 melanoma-bearing mice through transplantable animal tumor. Mice were treated with three doses of the polysaccharide LBPP (50, 100 and 200 mg/kg body weight) for 10 days. Tumor weight, relative spleen and thymus weight, lymphocyte proliferation, natural killer cell activity, delayed type hypersensitivity (DTH), phagocytic function of monocyte, serum hemolysis antibody and peripheral blood of tumor-bearing mice were studied. At the doses of 100 and 200 mg/kg, a significant decrease (P<0.01) in tumor formation, a significant increase (P<0.05) in relative spleen and thymus weight, natural killer cell activity, phagocytic function of monocyte, lymphocyte proliferation, and serum hemolysis antibody, and a significant improvement of peripheral blood abnormality (P<0.05) and anemia (P<0.01) were observed. Results of these studies demonstrated that the polysaccharide LBPP had anti-tumor activity, which was mediated by immunomodulation and leukogenic and antianemic actions.     

Low-dose curcumin leads to the inhibition of tumor growth via enhancing CTL-mediated antitumor immunity

Curcumin, a yellow pigment extracted from turmeric, is widely used to inhibit tumor progression. Since it can either promote or suppress the immune system, how curcumin affects the immune system in tumor-bearing bodies is not yet clear. Our study found that tumor-bearing mice treated consecutively once a day with low-dose curcumin for ten days led to a retarded tumor growth and a longer survival, which might be contributed to T cell-mediated adaptive immune response. The in vitro study also showed that a high-dose curcumin decreases T cells whereas a low-dose increases T cells derived from 3LL tumor-bearing mice, especially CD8+ T cells. Accordingly, these increased CD8+ T cells exhibited the enhancement of IFN-γ secretion, proliferation and cytotoxicity specifically against 3LL tumor cells, which may result in the success of antitumor immunity. Our research demonstrated a beneficial effect of curcumin on CD8+ T cells derived from tumor-bearing mice, which can provide a potential application in anti-tumor therapy.     

灵芝提取物抗肿瘤作用的实验研究

目的探讨一种灵芝提取物体内外抗肿瘤作用。方法应用小鼠H22肝癌移植瘤模型,研究灵芝提取物的体内抗肿瘤作用;采用MTT法检测灵芝提取物的体外抗肿瘤活性。结果①灵芝提取物对H22移植瘤呈剂量依赖性抑制作用,500 mg.kg-1组的瘤重抑制百分率达59.3%,与阴性对照组比较有统计学差异（P〈0.01）;②MTT法检测显示灵芝提取物对K562和HL60两种瘤株有较强抑制作用,对SMMC7221、HepG2、SW480、SW1116和SGC7901五种瘤株抑制作用较弱。结论灵芝提取物在体外及体内均有较强的抗肿瘤作用,其有效成分及机制有待进一步研究。     

Design of curcumin-loaded PLGA nanoparticles formulation with enhanced cellular uptake, and increased bioactivity in vitro and superior bioavailability in vivo

Curcumin, a yellow pigment present in the spice turmeric (Curcuma longa), has been linked with antioxidant, anti-inflammatory, antiproliferative, anticancer, antidiabetic, antirheumatic, and antiviral effects, but its optimum potential is limited by its lack of solubility in aqueous solvents and poor oral bioavailability. We employed a polymer-based nanoparticle approach to improve bioavailability. Curcumin was encapsulated with 97.5% efficiency in biodegradable nanoparticulate formulation based on poly (lactide-co-glycolide) (PLGA) and a stabilizer polyethylene glycol (PEG)-5000. Dynamic laser light scattering and transmission electron microscopy indicated a particle diameter of 80.9 nm. This curcumin, renamed from hereon “as curcumin (NP)”, was characterized for its biological activity. In vitro curcumin (NP) exhibited very rapid and more efficient cellular uptake than curcumin. Estrase staining revealed that curcumin (NP) was at least as potent as or more potent than curcumin in inducing apoptosis of leukemic cells and in suppressing proliferation of various tumor cell lines. When examined by electrophoretic gel shift mobility assay, curcumin (NP) was more active than curcumin in inhibiting TNF-induced NF-κB activation and in suppression of NF-κB-regulated proteins involved in cell proliferation (cyclin D1), invasion (MMP-9), and angiogenesis (VEGF). In mice, curcumin (NP) was more bioavailable and had a longer half-life than curcumin. Overall we demonstrate that curcumin-loaded PLGA nanoparticles formulation has enhanced cellular uptake, and increased bioactivity in vitro and superior bioavailability in vivo over curcumin.     

Characterization and anti-tumor activity of pollen polysaccharide

The polysaccharide LBPP was extracted and isolated from the pollen of Brassica napus L., and the anti-tumor activity was evaluated on Sarcoma 180-bearing mice and B16 melanoma-bearing mice through transplantable animal tumor. Mice were treated with three doses of the polysaccharide LBPP (50, 100 and 200 mg/kg body weight) for 10 days. Tumor weight, relative spleen and thymus weight, lymphocyte proliferation, natural killer cell activity, delayed type hypersensitivity (DTH), phagocytic function of monocyte, serum hemolysis antibody and peripheral blood of tumor-bearing mice were studied. At the doses of 100 and 200 mg/kg, a significant decrease (P<0.01) in tumor formation, a significant increase (P<0.05) in relative spleen and thymus weight, natural killer cell activity, phagocytic function of monocyte, lymphocyte proliferation, and serum hemolysis antibody, and a significant improvement of peripheral blood abnormality (P<0.05) and anemia (P<0.01) were observed. Results of these studies demonstrated that the polysaccharide LBPP had anti-tumor activity, which was mediated by immunomodulation and leukogenic and antianemic actions.     

Curcumin and its formulations: potential anti-cancer agents

Curcumin, one of the most studied chemopreventive agents, is a natural compound extracted from Curcuma longa L. Extensive research over the last half century has revealed that curcumin can inhibit the proliferation of various tumor cells in culture, prevent carcinogen induced cancers in rodents and inhibit the growth of human tumors in xenotransplant or orthotransplant animal models. Several phase I and phase II clinical trials indicated that curcumin is quite safe and may exhibit therapeutic efficacy. The utility of curcumin is limited by its lack of water solubility and relatively low in vivo bioavailability. Multiple approaches including nanoparticles, liposomes, micelles and phospholipid complexes are being sought to overcome these limitations. This review describes the general properties of curcumin and its potential effect against cancer including evidences of its antitumor action in vitro, in vivo, clinically and the strategies to overcome its low bioavailability.     

Anti-tumor effect of gallic acid on LL-2 lung cancer cells transplanted in mice.

We previously reported that gallic acid (3,4,5-trihydroxybenzoic acid), a naturally occurring plant phenol, can induce apoptosis in four kinds of human lung cancer cell lines in vitro. The present study further investigated the in vivo anti-tumor effects of orally administered gallic acid. Gallic acid reduced cell viability of LL-2 mouse lung cancer cells in vitro dose dependently, with a 50% inhibitory concentration (IC50) value of around 200 microM. C57Black mice were transplanted with LL-2 cells, and administered gallic acid (1 mg/ml in drinking water, ad libitum) and/or cisplatin (4 mg/kg i.p. injection, once a week). The average weight of the transplanted tumors, obtained at 29 days after transplantation, in the mice of control, gallic acid-treated cisplatin-treated and cisplatin plus gallic acid-treated groups was 4.02, 3.65, 3.19 and 1.72 g, respectively. The average tumor weight of the mice treated with cisplatin combined with gallic acid was significantly smaller than that of the control group (p<0.05). The amount of apoptotic cells in the tumor tissues of mice treated with gallic acid and/or cisplatin was significantly higher than those of the control mice. Combination of gallic acid and cisplatin increased the tumor cell apoptosis compared with the treatment with cisplatin alone. The present findings suggest that the combination of gallic acid with an anti-cancer drug, including cisplatin, may be an effective protocol for lung cancer therapy.     

姜黄素体内外逆转结肠癌的5-氟尿嘧啶耐药研究及机制探讨

摘要：目的 探究姜黄素在体内外对结肠癌5-氟尿嘧啶（5-FU）耐药的逆转作用及其相关机制。 方法 以SW480结肠癌细胞为亲代细胞,构建5-FU耐药细胞株SW480R,采用MTT法检测SW480R细胞的耐药指数以及不同浓度的姜黄素对SW480R细胞增殖能力的影响;流式细胞术检测姜黄素对SW480R细胞周期和凋亡的影响;Western blot检测姜黄素对SW480R细胞上皮-间充质转化（epithelial-mesenchymal transition, EMT）相关蛋白以及Wnt信号通路相关蛋白的影响;采用裸鼠移植瘤模型检测肿瘤体积变化情况,计算姜黄素的体内抑瘤率,Western blot检测肿瘤组织中相关蛋白变化。结果 SW480R的耐药指数为12.16,姜黄素能够剂量依赖性地抑制SW480R细胞的增殖,阻滞SW480R细胞周期于G0/G1期,以及诱导SW480R细胞凋亡;Western blot结果表明姜黄素能够在体内外抑制EMT和Wnt信号通路的活性;体内实验表明姜黄素能够有效抑制裸鼠移植瘤的生长。结论 姜黄素能够在体内外逆转结肠癌的5-FU耐药,其机制可能是通过调控Wnt信号通路从而抑制EMT的发生。     

Curcumin inhibits WEHI-3 leukemia cells in BALB/c mice in vivo

Curcumin (1, 7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5- dione), a natural polyphenol product of the plant Curcuma longa, exhibited potent inhibitory activities against proliferation, induced cell cycle arrest and exhibited the induction of apoptosis in several tumor cell lines. In our previous studies, we have shown that curcumin induced cell cycle arrest and apoptosis on human leukemia HL-60 and mouse leukemia WEHI-3 cells; there are no reports regarding whether or not it affects leukemia cells in vivo. In the present study, we investigated the effects of curcumin on WEHI-3 in BALB/c mice and the results indicated that curcumin reduces the percentage of Mac-3 marker, which is the precursor of macrophage. Curcumin induced significant effects on the population of B cells from murine leukemia in vivo. We also investigated the weights of spleen and liver from murine leukemia and the results showed that curcumin reduced the weight of the liver and spleen. From the pathological examinations, the effects of curcumin on the liver and spleen from mice after being injected with WEHI-3 cells were apparent. Both organs were enlarged. In conclusion, curcumin affect WEHI-3 cells in vivo.     

姜黄素通过调控miR-7641/PTPN14分子轴抑制乳腺癌发展进程的分子机制研究

目的 探讨姜黄素通过调控miR-7641/PTPN14分子轴抑制乳腺癌发展进程的分子机制。方法 采用实时荧光定量PCR（qRT-PCR）检测乳腺癌患者癌组织及细胞系中miR-7641表达情况;使用Kaplan-Meier方法作乳腺癌患者生存曲线;采用不同浓度的姜黄素处理细胞,或转染miR-7641 mimic、Anti-miR-7641及pcDNA-PTPN14载体,采用qRT-PCR检测miR-7641表达情况,MTT实验及克隆形成实验检测细胞增殖能力,Transwell小室法检测细胞迁移及侵袭,western blotting检测Ki67、pcDNA、CyclinD1、Bax、Bcl-2、caspase-3、caspase-8蛋白表达水平,采用双荧光素酶报告基因系统检测miR-7641与PTPN14靶向调控关系。结果 与癌旁组织或乳腺正常上皮细胞比较,miR-7641在乳腺癌患者癌组织及乳腺癌细胞系中高表达（P<0.01、0.001）,且miR-7641能够明显促进乳腺癌细胞的增殖、迁移及侵袭（P<0.05、0.01、0.001）,并促进Ki67、pcDNA、CyclinD1、Bcl-2蛋白表达,抑制Bax、caspase-3、caspase-8蛋白表达;miR-7641与PTPN14 3''-UTR靶向结合,姜黄素通过miR-7641/PTPN14分子轴抑制乳腺癌细胞的增殖、迁移及侵袭（P<0.01、0.001）,并抑制Ki67、pcDNA、CyclinD1、Bax蛋白表达,促进Bcl-2、caspase-3、caspase-8蛋白表达。结论 姜黄素可通过下调miR-7641促进PTPN14表达,进而抑制乳腺癌细胞的增殖、迁移及侵袭。     

蛇床子素-pH敏感型纳米粒的制备及其抗肿瘤活性的研究

目的:制备蛇床子素-Eudragit S100-pH敏感型纳米粒(Ost-S100-NP),并考察Ost-S100-NP体内和体外的抗肿瘤活性。方法:以乳化-溶剂扩散法制备Ost-S100-NP胶体溶液,体外采用MTT法考察Ost和Ost-S100-NP对宫颈癌细胞Hela-3的抑制作用;体内Ost对小鼠宫颈癌U14实体瘤的抗肿瘤试验采用常规的抗肿瘤试验方法,考察不同给药浓度、不同剂型对小鼠体质重、肿瘤生长和胸腺、脾及肝等脏器质量变化的影响。结果:Ost体外对Hela-3细胞的半数抑制浓度IC50分别为:61.25,56.87,48.46μg/mL;Ost-S100-NP的IC50则分别为46.57,40.23,37.46μg/mL;体内Ost及其Ost-S100-NP制剂对荷瘤小鼠的实体瘤抑制率最高可达40%,各给药组与空白对照组比较,差异有统计学意义(P<0.01)。结论:Ost-S100-NP体外和体内均有明显的抗肿瘤活性,且在治疗剂量下未出现毒性反应,有望开发成1种高效、低毒的Ost-S100-NP制剂。     

Sesamin exerts anti-tumor activity in esophageal squamous cell carcinoma via inhibition of TRIM44 and NF-κB signaling

Tripartite motif-containing 44 (TRIM44) is known to play an oncogenic role in multiple human cancers, including esophageal cancer. Sesamin possesses potent anti-inflammatory and anti-cancer properties for various cancers. This study is designed to unravel the biological functions of sesamin and TRIM44 in esophageal cancer. TRIM44 expression in esophageal squamous cell cancer (ESCC) cell lines and tissues was determined by RT-qPCR assay and Western blot. The effects of sesamin and TRIM44 on ESCC cell growth in vivo and in vitro were assessed by the mouse model and CCK-8 assay, respectively. We found that TRIM44 was significantly upregulated in ESCC cell lines and tissues when compared to their counterparts. Sesamin treatment or depletion of TRIM44 markedly reduced ESCC cell proliferation. The nuclear factor kappa B (NF-κB) and toll-like receptor 4 (TLR4) signaling pathway may be involved in sesamin-mediated TRIM44 suppression. Finally, we showed that oral administration of sesamin dramatically inhibited tumor growth or ESCC in nude mice. Our results suggest that sesamin exerts anti-tumor activity in ESCC via inhibition of NF-κB signaling pathway, demonstrating its potential for the treatment of esophageal cancer.