Antimetastatic Effects of Phyllanthus on Human Lung (A549) and Breast (MCF-7) Cancer Cell Lines

BACKGROUND: Current chemotherapeutic drugs kill cancer cells mainly by inducing apoptosis. However, they become ineffective once cancer cell has the ability to metastasize, hence the poor prognosis and high mortality rate. Therefore, the purpose of this study was to evaluate the antimetastatic potential of Phyllanthus (P. niruri, P. urinaria, P. watsonii, and P. amarus) on lung and breast carcinoma cells. METHODOLOGY/PRINCIPAL FINDINGS: Cytotoxicity of Phyllanthus plant extracts were first screened using the MTS reduction assay. They were shown to inhibit MCF-7 (breast carcinoma) and A549 (lung carcinoma) cells growth with IC(50) values ranging from 50-180 µg/ml and 65-470 µg/ml for methanolic and aqueous extracts respectively. In comparison, they have lower toxicity on normal cells with the cell viability percentage remaining above 50% when treated up to 1000 µg/ml for both extracts. After determining the non-toxic effective dose, several antimetastasis assays were carried out and Phyllanthus extracts were shown to effectively reduce invasion, migration, and adhesion of both MCF-7 and A549 cells in a dose-dependent manner, at concentrations ranging from 20-200 µg/ml for methanolic extracts and 50-500 µg/ml for aqueous extracts. This was followed by an evaluation of the possible modes of cell death that occurred along with the antimetastatic activity. Phyllanthus was shown to be capable of inducing apoptosis in conjunction with its antimetastastic action, with more than three fold increase of caspases-3 and -7, the presence of DNA-fragmentation and TUNEL-positive cells. The ability of Phyllanthus to exert antimetastatic activities is mostly associated to the presence of polyphenol compounds in its extracts. CONCLUSIONS/SIGNIFICANCE: The presence of polyphenol compounds in the Phyllanthus plant is critically important in the inhibition of the invasion, migration, and adhesion of cancer cells, along with the involvement of apoptosis induction. Hence, Phyllanthus could be a valuable candidate in the treatment of metastatic cancers

Growth Inhibition of Human Gynecologic and Colon Cancer Cells by Phyllanthus watsonii through Apoptosis Induction

Phyllanthus watsonii Airy Shaw is an endemic plant found in Peninsular Malaysia. Although there are numerous reports on the anti cancer properties of other Phyllanthus species, published information on the cytotoxicity of P. watsonii are very limited. The present study was carried out with bioassay-guided fractionation approach to evaluate the cytotoxicity and apoptosis induction capability of the P. watsonii extracts and fractions on human gynecologic (SKOV-3 and Ca Ski) and colon (HT-29) cancer cells. P. watsonii extracts exhibited strong cytotoxicity on all the cancer cells studied with IC50 values of ≤ 20.0 µg/mL. Hexane extract of P. watsonii was further subjected to bioassay-guided fractionation and yielded 10 fractions (PW-1→PW-10). PW-4→PW-8 portrayed stronger cytotoxic activity and was further subjected to bioassay-guided fractionation and resulted with 8 sub-fractions (PPWH-1→PPWH-8). PPWH-7 possessed greatest cytotoxicity (IC50 values ranged from 0.66 – 0.83 µg/mL) and was selective on the cancer cells studied. LC-MS/MS analysis of PPWH-7 revealed the presence of ellagic acid, geranic acid, glochidone, betulin, phyllanthin and sterol glucoside. Marked morphological changes, ladder-like appearance of DNA and increment in caspase-3 activity indicating apoptosis were clearly observed in both human gynecologic and colon cancer cells treated with P. watsonii especially with PPWH-7. The study also indicated that P. watsonii extracts arrested cell cycle at different growth phases in SKOV-3, Ca Ski and HT-29 cells. Cytotoxic and apoptotic potential of the endemic P. watsonii was investigated for the first time by bioassay-guided approach. These results demonstrated that P. watsonii selectively inhibits the growth of SKOV-3, Ca Ski and HT-29 cells through apoptosis induction and cell cycle modulation. Hence, P. watsonii has the potential to be further exploited for the discovery and development of new anti cancer drugs

Análise morfoanatômica comparativa da folha de Bauhinia forficata Link e B. variegata Linn. (Leguminosae, Caesalpinioideae) Comparative morphogical and anatomycal analyses of Bauhinia forficata Link and B. variegata Linn. (Leguminosae, Caesalpinioideae) leaves

Esse trabalho foi desenvolvido com o propósito de subsidiar o controle de qualidade das espécies medicinais Bauhinia forficata Link e B. variegata Linn. Essas foram coletadas em jardins e terrenos baldios de Curitiba, Paraná, Brasil. As amostras do limbo, pecíolo e pulvino foram fixadas em F.A.A. 50 e conservadas em etanol 70%. O seccionamento do material foi feito em micrótomo de rotação (material incluído em historresina) ou a mão livre e as lâminas foram montadas de forma permanente ou semipermanente. Foram realizadas análises em microscopia fotônica e eletrônica de varredura. A folha de B. forficata é alongada, com folíolos unidos em torno da metade do limbo, ápices agudos, superfície adaxial lisa e mucro entre os folíolos. Enquanto que, B. variegata possui folha mais larga que longa, com folíolos unidos na porção terminal do limbo e ápices obtusos. Nos pulvinos de B. variegata ocorrem camadas de colênquima, ausentes em B. forficata. O formato e a bainha de fibras dos pecíolos, em secção transversal, são distintos entre as espécies, assim como a distribuição dos estômatos e tricomas no limbo e a forma do bordo em secção transversal. B. variegata possui maior quantidade de idioblastos com drusas e compostos fenólicos. As características morfológicas e anatômicas de B. forficata e B. variegata contribuem para a identificação e diferenciação das espécies, sendo importantes para o controle de qualidade do material destinado à pesquisa e à indústria farmacêutica.<br>This work has been developed with the purpose of helping the quality control of the medicinal species Bauhinia forficata Link and B. variegata Linn., which were collected in gardens and vacant lots in Curitiba, Paraná, Brazil. The samples of leaf blade, petiole and pulvinus were fixed in FAA 50 and preserved in alcohol 70. The sectioning of the material was made in rotational microtome (with the material included in historesine) or manually and the blades were mounted in permanent or semipermanent way. Analyses in photonic and scanning electronic micros betscopy were performed. The leaf of B. forficata is elongated, with the leaflets united around the middle of the leaf blade, acute apices, smooth adaxial surface and mucroween the leaflets. The B. variegata, on the other hand, possesses a broader than long leaf, with the leaflets united in the terminal portion of the blade and obtuse apices. In the pulvinus of B. variegata there are collenchyma layers, which are absent in B. forficata. The shape and the petiole fibers sheath, in cross section, are distinct in both species, as well as the distribution of the stomata and trichomes in the leaf blade and the shape of the border in transversal section. B. variegata possesses a higher quantity of idioblasts with druses and phenolic compounds. The morphologic and anatomic characteristics of B. forficata an B. variegata contribute to the identification and distinction of the species and they are important for the control of the material destined to research and to the pharmaceutical industry