Bark beetles oxidize the defensive monoterpenes of their host trees both to detoxify them and to convert them into components of their pheromone system. This oxidation is catalysed by cytochrome P450 (CYP) enzymes and occurs in different stages of the insect. We identified two new CYP4 genes in the Chinese white pine beetle (Dendroctonus armandi), and carried out bioinformatic analysis one the full-length nucleic acid sequences and deduced amino acid sequences. Differential expression of the CYP4 genes was observed between sexes, and within these significant differences amongst development stages, fed on phloem of Pinus armandi and exposed to stimuli((±)- α-pinene, (R)-(+)- α-pinene, (S)-(-)-α-pinene, (S)-(-)-β-pinene and (+)-3-carene) at 8 and 24 h, and their interactions were found upon exposure to host monoterpenes. Increased expression of CYP4 genes suggested that they play a role in the detoxification of monoterpenes released by the host trees. The differential transcript accumulation patterns of these bark beetle CYP4 genes provides insight into the ecological interactions of D. armandi with its host pine.
The flight distance, flight time and individual flight activities of males and females of Dendroctonus armandi were recorded during 96-h flight trials using a flight mill system. The body weight, glucose, glycogen and lipid content of four treatments (naturally emerged, starved, phloem-fed and water-fed) were compared among pre-flight, post-flight and unflown controls. There was no significant difference between males and females in total flight distance and flight time in a given 24-h period. The flight distance and flight time of females showed a significant linear decline as the tethered flying continued, but the sustained flight ability of females was better than that of males. The females had higher glycogen and lipid content than the males; however, there was no significant difference between both sexes in glucose content. Water-feeding and phloem-feeding had significant effects on longevity, survival days and flight potential of D. armandi, which resulted in longer feeding days, poorer flight potential and lower energy substrate content. Our results demonstrate that flight distances in general do not differ between water-fed and starved individuals, whereas phloem-fed females and males fly better than water-fed and starved individuals.
Of all neoplasms found in women, cervical cancer has the third highest incidence and causes the fourth most deaths. All-trans retinoic acid (ATRA) may be with chemopreventive potential on cervical cancer, but the mechanisms underlying is not clear. To investigate the mechanisms, human cervical cancer HeLa cells were treated with ATRA for 1, 2, 3, or 4 days in vitro. We found that ATRA inhibited the growth of HeLa cells in a dose-dependent manner at the concentrations from 0.3 to 9.6 mumol/L. Flow cytometric analysis showed that HeLa cells were arrested at G0/G1 phase by ATRA, and the aneuploidy was found when cells were treated for 4 days, which is the first report that ATRA causes aneuploid cycle in HeLa cells. The expression of human telomerase catalytic subunit messenger RNA was decreased remarkably by ATRA. These findings suggested that the inhibition of telomerase activity and arrest of cells at G0/G1 phase might be the key steps through which ATRA inhibits the proliferation of HeLa cells. Our results provide a possible mechanistic explanation for the growth inhibitory effect of ATRA on HeLa cells. Therefore, retinoids may have therapeutic potential to complement current treatments of cervical cancers.
ABSTRACT. Anticancer activity of Bombyx batryticatus ethanol extract (BBE) against HeLa cells was studied using cell viability, DNA fragmentation, real-time polymerase chain reaction, and Western blot analyses. The BBE inhibited the growth and induced apoptosis of HeLa cells. The MTT assay indicated that the BBE induced cytotoxicity in HeLa cells in a time-and concentration-dependent manner. When HeLa cells were treated for 48 h, the 50% inhibitory concentration (IC 50 ) value for the BBE was 1.564 mg/mL. The microscopy results showed that HeLa cells were severely distorted and showed slow growth; some cells became round in shape when treated with 5 mg/mL BBE for 24 h. The DNA ladder results revealed excessive DNA fragmentation in HeLa cells treated with 7 mg/mL BBE for 36 h. The proapoptotic activity of the BBE was attributed to its ability to modulate the expression of Bcl-2 and Bax genes. The mRNA and protein expression levels of Bax were remarkably higher whereas those of Bcl-2 were lower than those in the control cells; this led to an increased Bax/Bcl-2 ratio in cells treated with the BBE for 36 h. The results suggest that the BBE might play an important role in tumor growth suppression by inducing apoptosis in human cervical cancer cells via the regulation of the Bcl-2-and Baxmediated apoptotic pathways.
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