In vitro cytotoxicity of curcuminoids against head and neck cancer HNO97 cell line / Citotoxicidade in vitro de curcuminoides contra a linhagem de células HNO97 de câncer de cabeça e pescoço

Oral squamous cell carcinoma (OSCC) is a malignant tumour of Head and Neck Cancer (HNC). The recent therapeutic approaches used to treat cancer have adverse side effects. The natural agents exhibiting anticancer activities are generally considered to have a robust therapeutic potential. Curcuminoids, one of the major active compounds of the turmeric herb, are used as a therapeutic agent for several diseases including cancer. In this study, the cytotoxicity of curcuminoids was investigated against OSCC cell line HNO97. Our data showed that curcuminoids significantly inhibits the proliferation of HNO97 in a time and dose-dependent manner (IC50=35 μM). Cell cycle analysis demonstrated that curcuminoids increased the percentage of G2/M phase cell populations in the treated groups. Treating HNO97 cells with curcuminoids led to cell shrinking and increased detached cells, which are the typical appearance of apoptotic cells. Moreover, flow cytometry analysis revealed that curcuminoids significantly induced apoptosis in a time-dependent manner. Furthermore, as a response to curcuminoids treatment, comet tails were formed in cell nuclei due to the induction of DNA damage. Curcuminoids treatment reduced the colony formation capacity of HNO97 cells and induced morphological changes. Overall, these findings demonstrate that curcuminoids can in vitro inhibit HNC proliferation and metastasis and induce apoptosis.

O carcinoma de células escamosas oral (OSCC) é um tumor maligno do câncer de cabeça e pescoço (HNC). As recentes abordagens terapêuticas usadas para tratar o câncer têm efeitos colaterais adversos. Os agentes naturais que exibem atividades anticâncer são geralmente considerados como tendo um potencial terapêutico robusto. Curcuminoides, um dos principais compostos ativos da erva cúrcuma, são usados como agente terapêutico para várias doenças, incluindo câncer. Neste estudo, a citotoxicidade dos curcuminoides foi investigada contra a linha de células OSCC HNO97. Nossos dados mostraram que os curcuminoides inibem significativamente a proliferação de HNO97 de forma dependente do tempo e da dose (IC50 = 35 μM). A análise do ciclo celular demonstrou que os curcuminoides aumentaram a porcentagem de populações de células da fase G2 / M nos grupos tratados. O tratamento das células HNO97 com curcuminoides levou ao encolhimento celular e ao aumento das células destacadas, que são a aparência típica das células apoptóticas. Além disso, a análise de citometria de fluxo revelou que os curcuminoides induziram significativamente a apoptose de uma maneira dependente do tempo. Além disso, em resposta ao tratamento com curcuminoides, caudas de cometa foram formadas nos núcleos das células devido à indução de danos ao DNA. O tratamento com curcuminoides reduziu a capacidade de formação de colônias das células HNO97 e induziu alterações morfológicas. No geral, esses achados demonstram que os curcuminoides podem inibir in vitro a proliferação e metástase de HNC e induzir apoptose.

Analysis of complete chloroplast genomes of Curcuma and the contribution to phylogeny and adaptive evolution.

Curcuma is an important member of Zingiberaceae. Many species of this genus are widely used in traditional medicine and have important cultural value in East Asia. Among them, C. longa is considered to be the main source of curcumin and has a very wide range of uses. The rapid development of molecular phylogeny has deepened our understanding of taxonomy and evolution of Curcuma. However, little is known about the chloroplast genome phylogeny and the genetic bases of adaptative evolution. In this work, we sequenced the complete chloroplast genome of 4 Curcuma species. Curcuma chloroplast genomes showed highly conserved structures and the length ranged from 159,423 bp to 152,723 bp. A total of 133 genes were observed. Multiple repeats and simple sequence repeats (SSRs) were detected. By comparing with related species, 7 highly variable regions were identified as potential specific DNA barcodes for species identification. Phylogenetic analysis of complete plastome sequences and specific data sets revealed discordance with expected genus boundary. Chloroplast phylogenetic relationships were better predicted by geography than by morphological and nuclear DNA, indicating a substantial existence of introgression. 9 genes were proved to have high posteriori probability in positive selection analysis, and 4 of them (psbA, psbD, PetA and rbcL) closely related to photosynthesis, implying that chloroplast genes may had undergone positive selection pressure in evolution. These results are of great significance for us to understand the genetic basis, phylogeny and adaptive evolution of Curcuma chloroplast.

Isolation of dihydrocurcuminoids from cell clumps and their distribution in various parts of turmeric (Curcuma longa).

In addition to well-known curcuminoids, three colored metabolites were isolated from cultured cell clumps that had been induced from buds on turmeric rhizomes. The isolated compounds were identified as dihydro derivatives of curcuminoids, dihydrocurcumin (dihydroCurc), dihydrodesmethoxycurcumin-a (dihydroDMC-a), and dihydrobisdesmethoxycurcumin (dihydroBDMC). The cell clumps did not contain dihydroDMC-b, an isomer of dihydroDMC-a. A comparison of the distribution profiles of curcuminoids and dihydrocurcuminoids in the cell clumps with those in the rhizomes, leaves, and roots revealed the following differences: Unlike rhizomes, the cell clumps, leaves, and roots contained dihydrocurcuminoids as the major colored constituents. Whereas dimethoxy compounds, curcumin and dihydrocurcumin, respectively, were most abundant in the rhizomes and leaves, one of the monomethoxy derivatives, dihydroDMC-a, was found most abundantly in the cell clumps and roots. While both dihydroDMC-a and b were detected in the rhizomes, dihydroDMC-b was not detectable in the cell clumps, leaves, or roots. The occurrence of only one of the two possible isomers of dihydroDMC suggests biosynthetic formation of dihydrocurcuminoids in turmeric.

[Establishment of high-yield suspension cell line of Curcuma zedoaria (Berg.) Rosc and study on the volatile oil synthesis-controlling with precursors].

The condition for high-yield suspension cell line and the precursors of volatile oil synthesis of Curcuma zedoaria (Berg.) Rosc were studied. The results showed that the light yellow particle callus was suitable for establishment of the high-yield suspension cell line. The optimum conditions for cell growth were MS medium added 15-30 g/L glucose and 15-30 g/L sucrose (1:1) as carbon source, the total concentration of 80 mmol/L nitrogen source combined NH4+ with NO3- (1:3), hormones of 3.0-5.0 mg/L 6-BA, 1.0 mg/L 2,4-D and dark culture after 10-15 days light culture. The 229 g/L cell (FW) and 2.11% content of volatile oil were obtained in vitro. The addition of precursors of calcium pantothenate, ammonium acetate and potassium acetate during the middle period of the cell suspension culture enhanced the volatile oil content respectively, and ammonium acetate was most effective among them. The highest yield of volatile oil obtained was 3.11% and 8.27 g/L respectively , which was 1.25 and 1.2 times of the control group.

[Histological and morphological studies on the rhizomes of Curcuma].

OBJECTIVE: To provide some new evidences for the identification of medicinal materials of Curcuma. METHOD: Microscopic observation was made to characterize the rhizomes of Curcuma. RESULT AND CONCLUSION: There were no obvious histological and morphological differences among the rhizomes of Curcuma. The distribution of oil cells and vascular bundles as well as the number and diameter of xylem vessels were considered to be the distinguishing features of their rhizomes.

[Pattern recognition of stereoscopic features of the leaves epidermis of medicinal Curcuma plants in China by image analysis].

OBJECTIVE: To provide new ways for classifying and identifying medicinal curcuma plants. METHOD: Based on classical taxonomy, the microscopic features of the epidermis of Curcuma plants in China were scored quantitatively by stereology and image analysis. RESULT: It showed that there were a lot of differences in density, size and shape of the epidermic cells among the medicinal Curcuma plants in China; that the average perimeter of the upper epidermic cells, average sectional area of the lower epidermis, stoma density and trichoma distribution of the upper and lower epidermis, etc. could be considered to be the main evidences for the microscopic identification of leaves of Curcuma. The 21 materials which belong to 11 species of Curcuma in China were systematized into 6 species, 1 species complex, 2 cultivated varieties. CONCLUSION: It can be concluded that stereology, and image analysis are advanced and feasible in pharmacognosy and taxonomy especially in the authentication of the relative and easily confused species.