A plant lignan, 3'-O-methyl-nordihydroguaiaretic acid, suppresses papillomavirus E6 protein function, stabilizes p53 protein, and induces apoptosis in cervical tumor cells.

Persistent infection with oncogenic human papillomaviruses (HPVs) is the most important factor in the induction of uterine cervical cancer, a leading cause of cancer mortality in women worldwide. Upon cell transformation, continual expression of the viral oncogenes is required to maintain the transformed phenotype. The viral E6 protein forms a ternary complex with the cellular E6-AP protein and p53 protein which promotes the rapid degradation of p53. Recent studies have revealed that lignans from the creosote bush (3'-O-methyl-nordihydroguaiaretic acid) can repress the viral promoter responsible for E6 gene expression. Work reported here shows that the lignan can subvert viral oncogene function resulting in stabilized p53 protein within treated HPV-containing tumor cells. The stabilized p53 is transcriptionally active as demonstrated by a luciferase reporter vector and induction of genes for Bax and PUMA proteins. Apoptosis is detected by annexin V binding to treated cells as analyzed by flow cytometry. Programmed cell death is confirmed by the induction of active caspases and TUNEL assay. Initiator caspase-9 is activated first, followed later by the effector caspase-3 enzyme. The stabilization and induced apoptosis are not observed within treated HPV-negative cervical tumor cells. Quantitative real time RT-PCR analysis of endogenous E6 gene transcription from the integrated HPV 16 promoter shows at least a fivefold repression of expression as compared to untreated cells. These results indicate that the loss of E6 protein in treated cells could be, in part, responsible for the stabilization of p53 within the lignan treated cells.

Cold acclimation strategy is highly variable among the sunfishes (Centrarchidae).

We tested the hypothesis that the physiological strategy for acclimating to low body temperature is similar among closely related fish. Largemouth bass (Micropterus salmoides), green sunfish (Lepomis cyanellus), bluegill sunfish (Lepomis macrochirus), black crappie (Pomonix nigromaculatus), and white crappie (Pomonix annularis), all members of the family Centrarchidae, were acclimated to 5 degrees and 25 degrees C. Morphometric variables (total mass, total length, organ masses) and enzyme activities (hexokinase; lactate dehydrogenase; and cytochrome oxidase in heart, liver, and muscle) were measured in 5 degrees C- and 25 degrees C-acclimated fish at 5 degrees and 25 degrees C assay temperatures. Each species displayed a distinct physiological response to cold acclimation that differed among tissues. These data suggest that the response to cold acclimation is highly variable within families. Our findings are consistent with other studies suggesting that acclimation responses are labile and may evolve independently even among closely related species.