Recent striking changes in histological differentiation and rate of human papillomavirus infection in squamous cell carcinoma of the lung in Okinawa, a subtropical island in southern Japan

Abstract

Aims: The incidence of lung cancer in Okinawa has been the highest in Japan since 1975, and squamous cell carcinoma (SCC), especially the well differentiated form, is the most prevalent form in Okinawa, although well differentiated SCC is relatively rare in mainland Japan. Furthermore, a high proportion of SCC of the lung in Okinawa was positive for human papillomavirus (HPV). In this study, we report recent striking changes in histological features and in the incidence of HPV infection.

Methods: In Okinawa between 1986 and 1998, 1109 surgically resected lung tumours were examined histopathologically. In addition, human papillomavirus infection was detected by the polymerase chain reaction and Southern blot analysis in SCC cases reported in 1993 and 1995-8. Non-isotopic in situ hybridisation of HPV DNA was also carried out.

Results: Up until 1994 SCC, especially the well differentiated form, was the most prevalent type of tumour. However, since 1995 the number of such cases has diminished steadily, accompanied by a slight rise in the incidence of adenocarcinoma. Although most present and past patients are heavy smokers, the incidence of SCC, especially the well differentiated form, continues to decrease steadily. Furthermore, in 1993, HPV was detected in 79% of all cases, and was particularly prevalent in the well differentiated form, but the rate fell to 68% in 1995, 35% in 1996, 23% in 1997, and 24% in 1998. The age distribution of patients, the male to female ratio, and the number of tumours overexpressing p53 protein did not change significantly over the study period, and thus did not correlate with changes in the differentiation of SCC.

Conclusions: The decreasing incidence of viral infection correlates strongly with the falling numbers of SCC cases, especially well differentiated cases. These findings suggest that HPV might be involved in the development of SCC of the lung, affecting the histological differentiation of SCC in particular, at least in Okinawa, a subtropical island in southern Japan.

Figure 1 (A) A representative example of well differentiated squamous cell carcinoma of the lung showing common histological features in such tumours in Okinawa (case detected in 1993). Note the presence of keratin pearls (indicated by a star). Haematoxylin and eosin staining; magnification, x150. (B) A representative example of poorly differentiated squamous cell carcinoma of the lung (case detected in 1998). Haematoxylin and eosin staining; magnification, x180.

Figure 2 Squamous cell carcinoma (SCC) of the lung: changes in the histological features. Open bars, well differentiated SCC; shaded bars, moderately differentiated SCC; black bars, poorly differentiated SCC.

Figure 3 Detection of HPV DNA in squamous cell carcinoma by non-isotopic in situ hybridisation. Note the presence of integrated (arrows) and episomal (arrowheads) forms of HPV in tumour cells (the same well differentiated squamous cell carcinoma case shown in fig 1). Keratin pearls indicated by an asterisk. Magnification, x250

Figure 4 Squamous cell carcinoma of the lung. Changes in the incidence of human papillomavirus positive cases.

Figure 5 Demonstration of human papillomavirus (HPV) DNA in the squamous cell carcinoma cases in 1998. Demonstration of the HPV-6 E6 region by the polymerase chain reaction and Southern blot analysis. The 189 bp band is shown in lane o (case 13 (1998) in table 5). The asterisk denotes the positive control (HPV-6 in plasmid pML); the star denotes the negative control (distilled water).

Figure 6 Demonstration of the human papillomavirus 11 (HPV-11) E6 region in the squamous cell carcinoma cases in 1998 by the polymerase chain reaction (PCR) and Southern blot analysis. Positive bands (230 bp) are shown in lanes a, d, and e (cases 1, 2, and 7 (1998) in table 5). The asterisk denotes the positive control (HPV-11 in plasmid pBR 322); the star denotes the negative control (distilled water).

Figure 7 (A and B) Demonstration of the human papillomavirus 16 (HPV-16) E6 region in the squamous cell carcinoma cases in 1998 by the polymerase chain reaction (PCR) and Southern blot analysis. No positive bands (240 bp) are shown. (C and D) Demonstration of the HPV-16 E7 region by PCR and Southern blot analysis. No positive bands (171 bp) are shown. The asterisk denotes the positive control (HPV-16 in plasmid pBR 322). The star denotes the negative control (distilled water).

Figure 8 (A and B) Demonstration of the human papillomavirus 18 (HPV-18) E6 region in the squamous cell carcinoma cases in 1998 by the polymerase chain reaction (PCR) and Southern blot analysis. No positive bands (160 bp) are demonstrated in the cases. (C and D) Demonstration of the HPV-18 E7 region by PCR and Southern blot analysis. A positive band (152 bp) is shown in lane s (case 5 (1998) in table 5). The asterisk denotes the positive control (HPV-18 in plasmid pBR 322); the star denotes the negative control (distilled water).