The predictive role of Bcl-2 expression on 6-year relapse-free and overall survival was mainly dependent on p53 expression.

Immunohistochemically detected p53 overexpression is an independent marker for shortened 6-year relapse-free and overall survival in node-negative patients with resectable breast cancers. Based on these findings, p53 overexpression should be used with other established prognostic factors, such as [3H]thymidine labeling index and estrogen receptor status, to further refine the prognostic assessment of node-negative breast cancer.

p53 expression appears to be indicative of clinical outcome in postmenopausal patients treated with tamoxifen. Whether p53 overexpression and weak bcl-2 expression are indicators of biologic aggressiveness, regardless of treatment, or of hormone resistance remains to be defined.

A causal link between human papillomavirus (HPV) infection and cervical cancer has been well established (2,12,14,23). A large number of HPV genotypes have been identified, and the mucosal HPV strains are divided into "high-risk" (HR) and "low-risk" (LR) categories on the basis of their association with cervical lesions. The HR types are more frequently found in premalignant or malignant lesions, LR types are found in benign lesions such as condylomata acuminata (18). Infection by HR HPV types has been demonstrated in almost 100% of cervical carcinoma (29), and it has been recently shown that persistent infection with the same genotype strongly increases the risk of developing high-grade preinvasive disease (11).The detection of HR HPV in cervical samples has been proposed to improve the efficacy of cervical carcinoma screening programs and to triage women with ambiguous or borderline cervical smears (1). Women with persistent HR HPV positivity have a clearly enhanced risk of developing a premalignant lesion and, hence, may be more closely monitored (3,15). Moreover, HR HPV testing may be associated with Pap smear to monitor women who have been treated for high-grade cervical intraepithelial neoplasia (CIN) (16,30). Several studies have revealed that HPV testing yields a high negative predictive value, approaching 100%, for high-grade CIN lesions and cervical carcinomas (lesions Ն CIN3) (21,22,31).Testing for HPV relies on the detection of viral DNA. The only test currently approved by the U.S. Food and Drug Administration for the detection of HPV DNA is the Hybrid Capture 2 (HC2) system (Digene Corporation, Gaithersburg, Md.). The HC2 assay is a ready-to-use test for routine diagnostics and uses a liquid hybridization format followed by signal amplification to detect 13 HR HPV types (i.e., genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68) by means of an RNA cocktail probe; the test does not distinguish individual HPV types. The sensitivity is approximately 4,700 viral copies/ml of cervical sample suspension. The evaluation of its laboratory performance has shown that HC2 is a reliable and reproducible test (4,6,8); both characteristics are fundamental for a test with potential widespread use. Different laboratories have also used many PCR-based methods. PCR methods are considered the "gold standard" for analytical sensitivity to detect infectious organisms, including HPV. However, PCRs for HPV detection are currently performed as "home brew" methods, lack standardization, show different sensitivity and specificity, are time-consuming, and require a demanding job for the laboratory. In the last years, some studies have compared the performance of HC2 and PCR, showing a good level of agreement between the two methods (19, 28).

Our study provides indirect evidence of a benefit from radiation therapy in preventing local breast cancer relapse, particularly among node-negative patients with tumors that express elevated levels of the p53 or GST-pi proteins or that express little or no Bcl-2 protein.

Interest in translational studies aimed at investigating the role of biologic markers in predicting clinical outcome of breast cancer patients and, in particular, response to specific treatments, has progressively increased. Among biologic variables presently under investigation, apoptosis markers, in particular Bcl-2 and Bax expression, are receiving much attention for their relationship with the cellular response to genotoxic damage in experimental tumors. Retrospective, independent studies were carried out by several research groups on about 5000 patients with breast cancer at different stages and with an adequate follow-up. The outcome of separate analyses as a function of treatment generally demonstrated that Bcl-2 overexpression, which correlates with biologic features of a differentiated phenotype (slow proliferation, high steroid receptor levels, absence of p53 and c-erB-2 expression), is associated with a favorable outcome. Such a finding is mainly evident following surgery as well as endocrine treatment. Conversely, no or weak Bcl-2 expression, alone or in association with bax overexpression, appears indicative of a radiation response, and preliminary emerging evidence supports the involvement of such an association of apoptosis-related markers even as predictors of long-term response to neoadjuvant cytotoxic treatment. Although the findings of an involvement of Bcl-2 and Bax as determinants of treatment response should be confirmed within the context of randomized clinical trials, they indicate a combined consideration of proteins that negatively and positively regulate apoptosis in translational studies on the effect of chemical and physical agents at a cellular level.

We investigated 27 pleomorphic carcinomas of the lung for exon 1 K-ras gene mutations using polymerase chain reaction-single-strand conformation polymophism analysis and direct sequencing. All pleomorphic carcinomas were biphasic, that is, composed of an adeno-, squamous-or large-cell-carcinomatous component associated with a spindle-and/or giant-cell component. Of 27 cases, six (22%) showed K-ras codon 12 mutations, which is a figure higher than that previously reported on in pure sarcoma-like pleomorphic carcinomas. Five tumors displayed the same mutation in both the epithelial and the sarcomatoid components, whereas in one tumor the mutation was restricted to the epithelial component. All mutations occurred in smokers, and were transversions, including GGT (glycine) to TGT (cysteine) change in two cases, to GCT (alanine) in two and to GTT (valine) in two. No significant relationships were found between the occurrence and type of mutations and patients' survival or any other clinicopathological variable, suggesting that K-ras mutations are early events in the development of these tumors. Our results indicate that most, though not all, biphasic pleomorphic carcinomas of the lung are monoclonal in origin, and that cigarette smoking may have a causative role in the development of K-ras alterations in these tumors, as all mutations are transversions.

In 187 node-negative breast cancers, the expression of laminin receptors and collagenase IV was directly related in 52% of cases, independently of pathological (tumor size and histology) and biological (estrogen receptors and proliferative activity) features. Moreover, the presence of laminin receptors and collagenase IV did not appear to influence tumor proliferative activity, evaluated as 3H-thymidine labelling index. In this case series, relapse-free survival and overall survival at 6 years were significantly affected by tumor size, hormone receptor status and proliferative activity. Conversely, high levels of laminin receptors and collagenase IV failed to influence relapse-free or overall survival, whereas they were strong indicators of local-regional diffusion of the disease.