Indole-3-carbinol and tamoxifen cooperate to arrest the cell cycle of MCF-7 human breast cancer cells.

The current options for treating breast cancer are limited to excision surgery, general chemotherapy, radiation therapy, and, in a minority of breast cancers that rely on estrogen for their growth, antiestrogen therapy. The naturally occurring chemical indole-3-carbinol (I3C), found in vegetables of the Brassica genus, is a promising anticancer agent that we have shown previously to induce a G1 cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. Combinations of I3C and the antiestrogen tamoxifen cooperate to inhibit the growth of the estrogen-dependent human MCF-7 breast cancer cell line more effectively than either agent alone. This more stringent growth arrest was demonstrated by a decrease in adherent and anchorage-independent growth, reduced DNA synthesis, and a shift into the G1 phase of the cell cycle. A combination of I3C and tamoxifen also caused a more pronounced decrease in cyclin-dependent kinase (CDK) 2-specific enzymatic activity than either compound alone but had no effect on CDK2 protein expression. Importantly, treatment with I3C and tamoxifen ablated expression of the phosphorylated retinoblastoma protein (Rb), an endogenous substrate for the G1 CDKs, whereas either agent alone only partially inhibited endogenous Rb phosphorylation. Several lines of evidence suggest that I3C works through a mechanism distinct from tamoxifen. I3C failed to compete with estrogen for estrogen receptor binding, and it specifically down-regulated the expression of CDK6. These results demonstrate that I3C and tamoxifen work through different signal transduction pathways to suppress the growth of human breast cancer cells and may, therefore, represent a potential combinatorial therapy for estrogen-responsive breast cancer.

Indole-3-carbinol inhibits the expression of cyclin-dependent kinase-6 and induces a G1 cell cycle arrest of human breast cancer cells independent of estrogen receptor signaling.

Indole-3-carbinol (I3C), a naturally occurring component of Brassica vegetables such as cabbage, broccoli, and Brussels sprouts, has been shown to reduce the incidence of spontaneous and carcinogen-induced mammary tumors. Treatment of cultured human MCF7 breast cancer cells with I3C reversibly suppresses the incorporation of [3H]thymidine without affecting cell viability or estrogen receptor (ER) responsiveness. Flow cytometry of propidium iodide-stained cells revealed that I3C induces a G1 cell cycle arrest. Concurrent with the I3C-induced growth inhibition, Northern blot and Western blot analyses demonstrated that I3C selectively abolished the expression of cyclin-dependent kinase 6 (CDK6) in a dose- and time-dependent manner. Furthermore, I3C inhibited the endogenous retinoblastoma protein phosphorylation and CDK6 phosphorylation of retinoblastoma in vitro to the same extent. After the MCF7 cells reached their maximal growth arrest, the levels of the p21 and p27 CDK inhibitors increased by 50%. The antiestrogen tamoxifen also suppressed MCF7 cell DNA synthesis but had no effect on CDK6 expression, while a combination of I3C and tamoxifen inhibited MCF7 cell growth more stringently than either agent alone. The I3C-mediated cell cycle arrest and repression of CDK6 production were also observed in estrogen receptor-deficient MDA-MB-231 human breast cancer cells, which demonstrates that this indole can suppress the growth of mammary tumor cells independent of estrogen receptor signaling. Thus, our observations have uncovered a previously undefined antiproliferative pathway for I3C that implicates CDK6 as a target for cell cycle control in human breast cancer cells. Moreover, our results establish for the first time that CDK6 gene expression can be inhibited in response to an extracellular antiproliferative signal.

National Delphi study to determine competencies for nursing leadership in public health.

PURPOSE: To identify competencies needed by nurse leaders in public health programs. DESIGN: Five-round national Delphi. SAMPLE: Convenience sample of members of major public health nursing associations and nurse and non-nurse public health leaders in the USA. METHODS: Mailed survey in 1994-1995 using a modified snowball technique based on a modification of the Pew Foundation health professions' competencies for Round 1. Four additional rounds produced consensus. FINDINGS: Initially, 62 competencies were identified. Factor analysis resulted in four factors: political competencies, business acumen, program leadership, and management capabilities; 57 competencies were clustered in the four groupings and accounted for 91.4% of the variance. CONCLUSIONS: Graduate schools in nursing and public health must prepare students with broad-based competencies from a variety of disciplines. Findings of this national survey provide a database for curriculum development and evaluation of programs to prepare nurse leaders for roles in public health-based delivery systems.

Observation evaluation to assess race and educational bias in state-mandated standard testing of nurse aides in nursing homes.

This article presents an assessment of whether race, education, gender, or other testing bias was present in a state-mandated nurse aide competency test. This assessment was carried out with data from two sources: (a) a statewide standardized test for all nurse aides that was given by a nationally known testing company, (b) an independent observational evaluation with a Behaviorally Anchored Rating Scale (BARS) for nurse aides' performance that was carried out by the investigators. The results show that race and education level were predictors of performance on written and manual portions of the standardized test. Gender, age, and years of experience were also shown to predict test success. Comparing data from the two sources suggests that a possible bias in the standardized nurse aid test. The independent observation of performance on the job with the BARS is shown to be less biased.

Molecular variants in the P450c11AS gene as determinants of aldosterone synthase activity in the Dahl rat model of hypertension.

Dahl salt-sensitive (S) and salt-resistant (R) rats are widely used to study genetic determinants of salt-sensitive hypertension. Differences in blood pressure under a high sodium diet in these two strains may be due to differences in the synthesis of 18-OH-11-deoxycorticosterone (18-OH DOC). This difference in 18-OH-DOC synthesis is due to mutations in the Dahl R rat's gene for P450c11 beta (11 beta-hydroxylase), an adrenal enzyme involved in the synthesis of both corticosterone and 18-OH DOC from 11-deoxycorticosterone. Aldosterone/renin ratios in plasma and in the adrenals are greater in Dahl S than R rats, suggesting an altered physiologic relationship between the renin-angiotensin and aldosterone systems between these strains. We demonstrate that the mRNA for P450c11AS, (aldosterone synthase), an enzyme required for aldosterone synthesis, is identical in the Dahl S rat and in normotensive Sprague-Dawley rats, but that P450c11AS mRNA from the Dahl R rat contains 7 mutations that result in two amino acid substitutions. These two changes result in a form of P450c11AS that has a greater apparent Vmax and lower apparent Km, resulting in an enzyme that catalyzes the conversion of 11-deoxycorticosterone to aldosterone at a greater rate in Dahl R rats than the P450c11AS in Dahl S rats or Sprague-Dawley rats. Although plasma and adrenal renin are lower in Dahl S versus R rats, the regulation of P450c11AS mRNA expression in rats fed a low and high salt diet are identical in these strains. The current findings may explain both the reduced aldosterone concentrations and increased aldosterone/renin ratios previously reported in the Dahl S versus Dahl R rat.