Flow cytometric dna analysis of hepatocellular carcinoma

Abstract

The prognostic value of nuclear DNA content was studied retrospectively using flow cytometry in 203 cases of resected hepatocellular carcinoma. The occurrence of DNA aneuploidy, which was detected in 50% of patients, correlated significantly with tumor size and the presence of vascular invasion or intrahepatic metastasis. Overall, patients with DNA aneuploid tumors had a significantly worse prognosis than those with DNA diploid tumors (P < 0.001) and, also in subdivided groups by tumor size (P < 0.01). Among DNA aneuploid patients, the survival times were significantly shorter for patients with a low DNA index (4.5) than for those with a high DNA index (21.5) (P < 0.05). In a Cox multivariate analysis, nuclear DNA content provided significant prognostic value (P = 0.008), as did vascular invasion (P = 0.001) and intrahepatic metastasis (P = 0.005). These results indicated that nuclear DNA content has an important prognostic value in hepatocellular carcinoma. Cancer 67:939-944,1991.EPATOCELLULAR CARCINOMA (HCC) iS one Of the H most frequent fatal tumors in Asia and Africa.' In Japan it is the third major cause of cancer death in males and the fifth among females.2 The 1-year survival rate is 55% even in resected Despite the argument that clinicopathologic factors affect the long-term survival rate,'y6 we often experienced an unsuspected clinical course or prognosis. The biological behavior of HCC is still unknown.Our goal was to develop a flow cytometric method for measuring the nuclear DNA content of HCC and to evaluate whether or not it had prognostic value as an independent factor. The relationship ofthe nuclear DNA content and other conventional prognostic variables was examined, and the survival data were analyzed using the Cox step-wise multivariate analysis' in a computerized program.From the First Department of Surgery, Hyogo College of Medicine, Nishinomiya, Japan.The authors thank Masahiro Kaji, Tomio Nakajima, and Hirotsugu Kubo for technical assistance.Address for reprints: Jiro Fujimoto, MD, First Department of Surgery, Hyogo College of Medicine, 1-1, Mukogawa-Cho, Nishinomiya 663, Japan.Accepted for publication August 20, 1990 Materials and Methods PatientsThe DNA ploidy analysis was done by flow cytometry from 203 patients receiving hepatectomy for hepatocellular carcinoma during the period from July 1973 to September 1987. The mean age of the patients was 57 years (range, 22-75), 88% of whom were male. These 203 patients received hepatectomy but did not undergo a palliative operation, such as hepatic artery ligation or cannulation. Chronic liver disease was noted as follows: cirrhosis, 168 (83%); and chronic hepatitis, fibrosis, or both, 20 (9.8%). Two samples taken from different places in the tumor were analyzed in 52 patients, whereas single samples were analyzed in 15 1 patients. The medical records of each patient were reviewed to confirm the sex, age, serum hepatitis B antigen (HBsAg), and serum alpha-fetoprotein (AFP). Pathologic examinationTumor size was the largest diameter recorded in...

Genetics of hepatocellular tumors

“…Genetics of hepatocellular tumors P Laurent-Puig and J Zucman-Rossi in tumor cells suggesting a global gain of genetic material, in half of the analysed HCC cases (Ezaki et al, 1988;Fujimoto et al, 1991;Chiu et al, 1992). Hyperploidy is also found in 43% of dysplastic lesions observed in cirrhotic disease (Thomas et al, 1992).…”

Heterogeneity of DNA content in multiple synchronous hepatocellular carcinomas

“…It has been shown that DNA content analysis is an important predictor of prognosis, which is better in diploid than in aneuploid tumours (Fujimoto et al, 1991;Chiu et al, 1992), although some authors have suggested that the advanced tumour stage of large lesions may negate the influence of DNA ploidy status on patient outcome (McEntee et al, 1992). These studies were limited to analysis of a single tumour nodule in patients with HCC.…”

“…Quantitative DNA analysis reflects the total chromosomal content of tumour cells. In terms of the cell nuclear DNA content of HCC, it has been reported (Fujimoto et al, 1991;Chiu et al, 1992) that an aneuploid DNA pattern indicates poor prognosis. With regard to the heterogeneity of the DNA ploidy pattern, HCC has been reported to be mostly homogeneous within the same tumour (Kuo et al, 1987;Nagasue et al, 1993;Ng et al, 1994).…”

Intratumoral Heterogeneity of DNA Ploidy and Regional Differences in Epidermal Growth Factor and Epidermal Growth Factor Receptor of Esophageal Carcinoma.

“…Furthermore, the autocrine mechanism of EGF and EGF-R could be one of the contributory factors on the development of DNA abnormalities during tumor proliferation. intratumoral heterogeneity; cytofluorometry; immunohistochemistry; esophageal carcinoma With the aid of flow cytometry and cytofluorometry, the determination of the DNA content in various types of tumors (Sannohe and Hiratsuka 1981;Sugimachi et al 1986;Heikki et al 1990;Sing et al 1990;Fujimoto et al 1991) has been widely carried out to evaluate the degree of malignancy and prognosis. Compared with the flow cytometry, the cytofluorometry has been reported to have more advantage to measure the DNA content in specific target cells and to provide more accurate results from small tissue samples (Cusic et al 1990).…”

“…Although enough number of examined cases were available in this study, the present results seem to be compatible with the idea that multiple DNA types develop during carcinogensis (Rabinovitch et al 1988;Mckinley et al 1987). Furthermore, we speculate that the cancer cells with the most adaptable DNA stemline(s) grow faster, and eventually form a tumor Fujimoto et al 1991) have indicated that patients with aneuploidy have poorer prognosis than those with diploidy, whereas little information is available concerning the relationship between prognosis and DNA heterogeneity. Since patients with the DNA heterogeneity always have aneuploidy, it is reasonable to speculate that multiple aneuploidy may be present in such patients and that the prognosis of patients with DNA heterogeneity may be poorer than those with DNA homogeneity.…”