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Factors affecting Agrobacterium-mediated transformation in Hybanthus enneaspermus (L.) F. Muell
Ganeshan Sivanandhan,Chinnathambi Arunachalam,Venkatachalam Vasudevan,Gnanajothi Kapildev,Ali Alharbi Sulaiman,Natesan Selvaraj,Andy Ganapathi,임용표 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.2
Agrobacterium tumefaciens-mediated transformation system was established for Hybanthus enneaspermus using leaf explants with the strain LBA4404 harbouring pCAMBIA 2301 carrying the nptII and gusA genes. Sensitivity of leaf explants to kanamycin was standardized (100 mg/l) for screening the transgenic plants. Transformation parameters (OD, virulence inducer, infection time, co-cultivation period, bactericidal antibiotics, etc.) influencing the gene transfer and integration were assessed in the present investigation. Fourteen-day precultured explants were subjected with Agrobacterium strain LBA4404. Optimized parameters such as culture density of 0.5 OD600, infection time of 6 min, AS concentration of 150 lM with 3 days co-cultivation revealed maximum transformation efficiency based on GUS expression assay. The presence of gusA in transgenics was confirmed by polymerase chain reaction and Southern blotting analysis. The present transformation experiment yielded 20 shoots/explant with higher transformation efficiency (28 %). The protocol could be used to introduce genes for trait improvement as well as for altering metabolic pathway for secondary metabolites production.
Li, Feng,Zhang, Jingwen,Arfuso, Frank,Chinnathambi, Arunachalam,Zayed, M E,Alharbi, Sulaiman Ali,Kumar, Alan Prem,Ahn, Kwang Seok,Sethi, Gautam Springer-Verlag 2015 Archives of toxicology Vol.89 No.5
<P>The transcription factor nuclear factor kappa B (NF-kappa B) has attracted increasing attention in the field of cancer research from last few decades. Aberrant activation of this transcription factor is frequently encountered in a variety of solid tumors and hematological malignancies. NF-kappa B family members and their regulated genes have been linked to malignant transformation, tumor cell proliferation, survival, angiogenesis, invasion/metastasis, and therapeutic resistance. In this review, we highlight the diverse molecular mechanism(s) by which the NF-kappa B pathway is constitutively activated in different types of human cancers, and the potential role of various oncogenic genes regulated by this transcription factor in cancer development and progression. Additionally, various pharmacological approaches employed to target the deregulated NF-kappa B signaling pathway, and their possible therapeutic potential in cancer therapy is also discussed briefly.</P>
Lee, Hanwool,Baek, Seung Ho,Lee, Jong Hyun,Kim, Chulwon,Ko, Jeong-Hyeon,Lee, Seok-Geun,Chinnathambi, Arunachalam,Alharbi, Sulaiman Ali,Yang, Woong Mo,Um, Jae-Young,Sethi, Gautam,Ahn, Kwang Seok MDPI 2017 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.18 No.5
<P>Isorhynchophylline (Rhy) is an active pharmacological component of <I>Uncaria rhynchophylla</I> that has been reported previously to exert significant antihypertensive and neuroprotective effects. However, very little is known about its potential anti-cancer activities. This study was carried out to evaluate the anticancer effects of Rhy against various human carcinoma cell lines. We found that Rhy exhibited substantial cytotoxic effect against human hepatocellular carcinoma HepG2 cells when compared with other human carcinoma cell lines including those of lung, pancreas, prostate, head and neck, breast, multiple myeloma, brain and renal cell carcinoma. Rhy induced apoptosis as characterized by accumulation of cells in sub G1 phase; positive Annexin V binding; activation of caspase-8, -9, and -3; and cleavage of PARP (poly-ADP ribose polymerase). This effect of Rhy correlated with the down-regulation of various proteins that mediated cell proliferation, cell survival, metastasis, and angiogenesis. Moreover, cell proliferation, migration, and constitutive CXCR4 (C-X-C chemokine receptor type 4), MMP-9 (Matrix metallopeptidase-9), and MMP-2 expression were inhibited upon Rhy treatment. We further investigated the effect of Rhy on the oncogenic cell signaling cascades through phospho-kinase array profiling assay. Rhy was found to abrogate phospho-p38, ERK, JNK, CREB, c-Jun, Akt, and STAT3 signals, but interestingly enhanced phospho-p53 signal. Overall, our results indicate, for the first time, that Rhy could exert anticancer and anti-metastatic effects through regulation of multiple signaling cascades in hepatocellular carcinoma cells.</P>