Morindone from morinda citrifolia as a potential antiproliferative agent against colorectal cancer cell lines

There is an increasing demand in developing new, effective, and affordable anti-cancer against colon and rectal. In this study, our aim is to identify the potential anthraquinone compounds from the root bark of Morinda citrifolia to be tested in vitro against colorectal cancer cell lines. Eight potential anthraquinone compounds were successfully isolated, purified and tested for both in-silico and in-vitro analyses. Based on the in-silico prediction, two anthraquinones, morindone and rubiadin, exhibit a comparable binding affinity towards multitargets of β-catenin, MDM2-p53 and KRAS. Subsequently, we constructed a 2D interaction analysis based on the above results and it suggests that the predicted anthraquinones from Morinda citrifolia offer an attractive starting point for potential antiproliferative agents against colorectal cancer. In vitro analyses further indicated that morindone and damnacanthal have significant cytotoxicity effect and selectivity activity against colorectal cancer cell lines

Morindone from Morinda citrifolia as a potential antiproliferative agent against colorectal cancer cell lines

There is an increasing demand in developing new, effective, and affordable anti-cancer against colon and rectal. In this study, our aim is to identify the potential anthraquinone compounds from the root bark of Morinda citrifolia to be tested in vitro against colorectal cancer cell lines. Eight potential anthraquinone compounds were successfully isolated, purified and tested for both in-silico and in-vitro analyses. Based on the in-silico prediction, two anthraquinones, morindone and rubiadin, exhibit a comparable binding affinity towards multitargets of β-catenin, MDM2-p53 and KRAS. Subsequently, we constructed a 2D interaction analysis based on the above results and it suggests that the predicted anthraquinones from Morinda citrifolia offer an attractive starting point for potential antiproliferative agents against colorectal cancer. In vitro analyses further indicated that morindone and damnacanthal have significant cytotoxicity effect and selectivity activity against colorectal cancer cell lines

Disruption of mammalian dream complexes by human papilomavirus E7 proteins / Nurshamimi Nor Rashid

Human papillomaviruses have been identified as the major aetiological factor in cervical cancer. Constitutive expression of the high risk HPV E6 and E7 oncoproteins are important for malignant transformation in infected keratinocytes. In particular, E6 and E7 bind to and inactivate the cellular tumor suppressors p53 and pRB, respectively, thus delaying differentiation and inducing proliferation in suprabasal keratinocytes to enable HPV replication. One member of the pRB family, p130 appears to be an important target for E7 in promoting S-phase entry. Recently, it has been discovered that p130 is part of a large protein complex termed DREAM. The composition of DREAM is temporally regulated during the cell cycle; being associated with E2F-4 and either p107 or p130 in G0/G1 and with the B-myb transcription factor in S/G2. In this study, we addressed whether p130/DREAM complex is disrupted in HPVtransformed cell lines and whether this property is important for E6 and E7 function. We found that, p130-DREAM complex diminished in HPV-transformed cell lines (CaSki and SiHa) compared to control cell lines. However, p130/DREAM complex was reformed and cell cycle was arrested in HPV-transformed cell lines when E6/E7 expression was targeted by specific small hairpin RNAs. We further demonstrated that the profound G1 arrest in CaSki cells was dependent on p130/DREAM reformation by also targeting the expression of the DREAM component Lin-54 and p130. Moreover, p130/DREAM complex was completely disrupted in high risk HPV compared to low risk and cutaneous HPV when various types of HPV were expressed ectopically in T98G cells. Interestingly, one type of the cutaneous HPV, 48 has an ability to disrupt p130/DREAM quite dramatically although it binds to pocket protein with iv extremely low affinity. Furthermore we found that B-myb/DREAM complex is not critical in regulating S/G2 phase in CaSki cells as determined by G2/M-phase genes in real time PCR. Finally, we showed the ability of E7 in disrupting p130/DREAM complex through multiple mechanisms. Four different types of p130 mutants were expressed in pMSCV puro vector and transfected into CaSki and T98G cell lines. The results demonstrate that 16E7 must bind to p130 in order to induce the S/G2 phase in HPV-transformed cell line. Furthermore, the cutaneous HPV type (48E7) promotes the S phase by binding towards p21, the CDK inihibitor. The results show that continued HPV16 E6/E7 expression is necessary in cervical cancer cells to prevent cell-cycle arrest by a repressive p130/DREAM complex

A Preliminary Study in Search of Potential Peptide Candidates for a Combinational Therapy with Cancer Chemotherapy Drug

Cancer which caused by the growth and spreading of abnormal cells in an uncontrolled manner remains a major cause of death affecting millions of people. The current cancer chemotherapy treatment modalities have several disadvantages, mostly related to their undesirable side effects. In this study, we explore the potencies of selected cell penetrating antimicrobial peptides in combination with the widely used chemotherapy drug, Doxorubicin (DOX), to increase the specificity of anticancer chemotherapy. Screening the potential peptide candidates to be developed into chemotherapy drug combination led to identification of two most potent peptides, Tachyplesin 1 (TCH) and Latarcin 1 (LTC). Cell viability of normal liver cells was reduced to 50% by 20 µM of TCH or LTC, while liver cancer cell lines lost 50% of their viability at approximately 4 µM of these peptides. The combination of DOX with TCH peptide showed the higher levels of lactate dehydrogenase (LDH) leakage from cancer cells (80%) compared to normal cells (30%). The combinational treatment DOX-peptide showed significant (P < 0.01) increase in caspase 3/7 activity compared to DOX alone. Pre-treatment of the cells with TCH peptides prior to DOX treatment considerably increased the Caspase 3/7 activities in both cell types with significant increase (P < 0.05) in cancer cells compared to normal cells. Our study demonstrate that TCH peptide is a potential anticancer peptide that could be used in combinational therapy with cancer chemotherapy drugs. © 2017, Springer Science+Business Media, LLC, part of Springer Nature

Endoplasmic reticulum: a focal point of Zika virus infection

Zika virus (ZIKV) belongs to the Flavivirus genus of the Flaviviridae family. It is an arbovirus that can cause congenital abnormalities and is sexually transmissible. A series of outbreaks accompanied by unexpected severe clinical complications have captured medical attention to further characterize the clinical features of congenital ZIKV syndrome and its underlying pathophysiological mechanisms. Endoplasmic reticulum (ER) and ER-related proteins are essential in ZIKV genome replication. This review highlights the subcellular localization of ZIKV to the ER and ZIKV modulation on the architecture of the ER. This review also discusses ZIKV interaction with ER proteins such as signal peptidase complex subunit 1 (SPCS1), ER membrane complex (EMC) subunits, and ER translocon for viral replication. Furthermore, the review covers several important resulting effects of ZIKV infection to the ER and cellular processes including ER stress, reticulophagy, and paraptosis-like death. Pharmacological targeting of ZIKV-affected ER-resident proteins and ER-associated components demonstrate promising signs of combating ZIKV infection and rescuing host organisms from severe neurologic sequelae

Additional file 2: Figure S2. of HPV 16E7 and 48E7 proteins use different mechanisms to target p130 to overcome cell cycle block

The p130mE7 mutant is unable to bind 16E7 and HPV48 E7 binds to p21. (A) Nuclear extracts were prepared from T98G cells transfected with the pMSCVpuro vector encoding HA-tagged p130wt or p130mE7 and from cells transfected with the empty vector (con). Nuclear extracts (150 μg) were incubated with 20 μg 16E7 protein bound to glutathione-Sepaharose beads and the selected proteins were eluted and run on a western blot alongside inputs comprising 15 μg of each nuclear extract. The p130wt and p130mE7 were detected using an HA antibody probe. In addition to the p130 proteins, a non-specific (n.s.) band was seen with the input samples. (B) A GST binding assay was carried out using GST-tagged-E7- HA proteins and IVT [35S]-labelled p21. 25 μg of each GST16-E7-HA and GST48-E7-HA proteins were resolved by SDS-PAGE alongside inputs comprising 15 μg of nuclear extract and were detected using an HA antibody probe. Any bound p21 was visualised by autoradiography. The image is representative of three independent experiments. (JPG 35 kb

Additional file 1: Figure S1. of HPV 16E7 and 48E7 proteins use different mechanisms to target p130 to overcome cell cycle block

Expression of various p130 mutants in C33a and CaSki cell lines. pMSCV puro (Clontech, Mountain View, CA) constructed with p130 wt and p130 mutants (p130mE7, p130PM22 and p130 mE7/PM22) were ‘FuGENE 6’ transfected in C33a and CaSki cells. The T98G cell lines (HPV 16 E7 negative cell line) were used as a control cells. Transfected cells were puromycin selected and nuclear lysates were harvested 48 hours post transfection. Nuclear lysates were separated on a 10 % SDS-PAGE gel and western blotted onto a nitrocellulose membrane. (a) Endogenous p130 were detected by p130 (Santa Cruz) and β-actin (Sigma Aldrich) was used as a loading control. (b) Ectopically expressed p130 were detected by p130 (Santa Cruz) and HA (Roche) antibodies, respectively. β-actin (Sigma Aldrich) was used as a loading control. The image is representative of three independent experiments. (JPG 51 kb

The ABA392/pET30a protein of Pasteurella multocida provoked mucosal immunity against HS disease in a rat model

Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among bovines. The disease is caused by the Pasteurella multocida serotype B:2 bacteria. P. multocida B:2 has high mortality and morbidity rates and is spread through the intranasal and oral routes in bovines. In this study, our aim was to investigate the efficacy of the recombinant protein vaccine, ABA392/pET30a via intranasal inoculation by targeting the mucosal immunity. The constructed recombinant protein vaccine ABA392/pET30a was subjected to an animal study using Sprague Dawley rats. The study was divided into two parts: active and passive immunization studies. Both studies were carried out through the determination of immunogenicity (using Total White Blood Cell (TWBC) Count with Indirect ELISA) and histopathogenicity, analyzing (Bronchus Associated Lymphoid Tissue (BALT) formation) in lungs. As a result, the IgA and IgG development of both tested groups: group 1 (50μg/mL protein vaccine) and group 2 (100μg/mL protein vaccine) showed equivalent with the positive control group 4 (formalin-killed P. multocida B:2). However, there was a significant difference when compared with the negative control group 3 (normal saline). These results demonstrate that both the protein vaccine at the concentration 50μg/mL and 100μg/mL have the same efficacy as the commercially available positive control vaccine. From the studies, higher concentration of protein vaccine at 100μg/mL showed higher development of both IgA and IgG compared to 50μg/mL protein vaccine. Higher and rapid development of IgA compared to IgG showed that mucosal immunity has been induced through the intranasal administration of the protein vaccine. In addition, leucocytosis was observed at each dose of vaccination showed that the protein vaccine is capable to induce the immune responses of the host. Histopathogenicity studies of the vaccinated groups showed more BALT formation and no severe lesions after challenge compared to the negative control group. Besides, no inflammatory onsite or anaphylactic responses were observed after the intranasal inoculation which proved to be safer and provided longer lasting immunity. Therefore, recombinant protein vaccine ABA392/pET30a could be a potential candidate for intranasal administration which can provoke mucosal immunity against HS disease