Reiko Hirose

Biologic implications of the expression of vascular endothelial growth factor subtypes in ovarian carcinoma

Vascular endothelial growth factor (VEGF) has been identified as an important factor for tumor angiogenesis, which is essential for the growth, invasion, and metastasis of solid tumors. This study examines the clinical significance of VEGF subtypes in ovarian carcinoma.

Expression of vascular endothelial growth factor (VEGF) and its mRNA in uterine cervical cancers

SummaryTo know the potential of growth, invasion and metastasis of uterine cervical cancer associated with neovascularization, localization of vascular endothelial growth factor (VEGF) and microvessel density in tumours were determined by immunohistochemical staining, the levels of VEGF subtypes were determined by Western blot analysis and by a sandwich enzyme immunoassay, and the levels of VEGF subtype mRNAs were determined by reverse transcription polymerase chain reaction (RT-PCR) – Southern blot analysis in uterine cervical cancers. The relation between VEGF subtype expressions and microvessel density, histological types and clinical stages of uterine cervical cancers was analysed. The expression of VEGF was seen dominantly in the cancer cells, and correlated with microvessel density in uterine cervical cancers. Among the four subtypes of VEGF, the populations of VEGF165 and VEGF121 were dominant in normal uterine cervices and uterine cervical cancers. The levels of VEGF and VEGF165 and VEGF121 mRNAs were remarkably higher in some stage II and III/IV adenocarcinomas of the cervix than in other cases, including normal cervices. Therefore, the elevation of VEGF165 and VEGF121 might contribute to the relatively late advancing via angiogenic activity in some adenocarcinomas of the cervix.

Expressions of vascular endothelial growth factor (VEGF) and its mRNA in uterine endometrial cancers

To know the potential of growth, invasion and metastasis of uterine endometrial cancer associated with neovascularization, the expressions of VEGF and its mRNA, especially their subtypes, in uterine endometrial cancers and normal uterine endometria as controls were determined by Western blot analyses with a sandwich enzyme immunoassay and RT-PCR-Southern blot analysis, respectively, and the relation between their expressions and histological grades, grades of myometrial invasion and clinical stages of uterine endometrial cancers was analyzed. The levels of VEGF (VEGF165 and VEGF121) protein and mRNA were in a wide range and higher in normal uterine endometria than in the malignant counterparts. The levels of VEGF protein were higher in order of histopathological differentiation (normal uterine endometrium > well-differentiated (G1) > moderately differentiated (G2) and poorly differentiated (G3)) and those of VEGF protein and VEGF121 mRNA were lower in order of the advance of clinical stages (normal uterine endometrium > stage I > stage II > stages III and IV). There was, however, no significant difference in their levels among uterine endometrial cancers classified according to grades of myometrial invasion. This suggests that VEGF is downregulated during uterine endometrial cancer progression with dedifferentiation. Namely, VEGF in some endometrial cancers might contribute to the early process of advancing of malignancy via angiogenic activity.

Clinical implication of expression of progesterone receptor form A and B mRNAs in secondary spreading of gynecologic cancers

This study was designed to determine the clinical implication of expression of progesterone receptor form A (PR-A) and B (PR-B) mRNAs in secondary spreading of gynecologic cancers. Approximately equal expression of PR-A and PR-B mRNAs was designated as type AB and dominant expression of PR-B mRNA as type B. Alteration from type AB to type B in the metastatic cancers occurred in 3/8 cases of uterine endometrial cancers, 2/8 cases of uterine cervical cancers, and 2/8 cases of ovarian cancers. Other cancers revealed type B regardless of primary or metastatic status. Thus, all metastatic cancers studied revealed type B. These results suggest that transcription of PR-A mRNA may be damaged, which might lead to uncontrolled overexpression of PR-B mRNA in metastatic lesion, and that the type B status could reveal a highly malignant phenotype in these three gynecologic cancers.

Expression of platelet-derived endothelial cell growth factor (PD-ECGF) and its mRNA in uterine cervical cancers.

SummaryAngiogenesis contributes to the growth and secondary spreading of solid tumours. Platelet-derived endothelial cell growth factor (PD-ECGF) is identified as such an angiogenic factor. In the present study, the prognosis of the patients with high PD-ECGF uterine cervical cancers was worse than those with low PD-ECGF cancers, and PD-ECGF expression correlated with cellular proliferation and with vascular density and venous invasion in uterine cervical cancers. Therefore, PD-ECGF might contribute to the growth of uterine cervical cancers via angiogenesis related to vascular spreading. Furthermore, PD-ECGF and its mRNA had a wide range and were highly expressed in uterine cervical cancers, especially squamous cell carcinoma, regardless of clinical stage. Therefore, PD-ECGF in uterine cervical cancers might play a role of basic angiogenesis in all processes of advancing of uterine cervical cancers. This indicates that 5′-deoxy-5-fluorouridine might be highly effective in squamous cell carcinoma of the cervix, which possesses a high activity of thymidine phosphorylase to convert 5′-deoxy-5-fluorouridine to 5-fluorouracil, and that some angiogenic inhibitors of new capillary formation might be effective in the inhibition of tumour growth and spreading associated with angiogenesis.

Expression of basic fibroblast growth factor and its mRNA in advanced uterine cervical cancers

To know the potential of growth, invasion and metastasis of uterine cervical cancer cells associated with neovascularization, the expression of basic fibroblast growth factor (FGF) and its mRNA in uterine cervical cancers and normal uterine cervices as controls were determined by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction-Southern blot (RT-PCR-SB), respectively. Then, the relations between the expression and the histological grading and clinical staging in cervical cancers were analyzed. The levels of basic FGF and its mRNA were significantly higher in advanced primary uterine cervical cancers, regardless of histological type. Therefore, this status might contribute to the acceleration of growth, invasion, and metastasis with neovascularization in advanced uterine cervical cancers.

Progestins suppress estrogen-induced expression of vascular endothelial growth factor (VEGF) subtypes in uterine endometrial cancer cells

Vascular endothelial growth factor (VEGF) contributes to the early advancement of uterine endometrial cancers that conserve hormone dependency via angiogenic activity. This process prompted us to study sex steroidal suppression of VEGF expression in Ishikawa cells (a line of well-differentiated uterine endometrial cancer cells). Estrogen transiently induced VEGF subtype (VEGF165 and VEGF121) secretion from Ishikawa cells. Progestins (progesterone, medroxyprogesterone acetate (MPA) and 17 alpha-hydroxyprogesterone) suppressed the estrogen-induced events. In conclusion, progestins could suppress VEGF-related angiogenic potential, which contributes to tumor growth in the early stage of uterine endometrial cancers that conserve estrogen dependency.

Expression of platelet-derived endothelial cell growth factor (PD-ECGF) and its mRNA in uterine endometrial cancers

To determine the potential of growth, invasion and metastasis of uterine endometrial cancer cells associated with neovascularization, the expressions of platelet-derived endothelial cell growth factor (PD-ECGF) and its mRNA in uterine endometrial cancers and in normal uterine endometria as controls were determined and the relationship between their expressions and histological grades, grades of myometrial invasion and clinical stages of uterine endometrial cancers was analyzed. The levels of PD-ECGF were significantly higher in uterine endometrial cancers of well-differentiated grade (G1) with invasion to < or =1/2 myometrium (B) and of stage 1 than in those of moderately and poorly differentiated grades (G2 and G3, respectively) limited to endometrium (A) and with invasion to >1/2 myometrium (C) and of stages II and III/IV and in normal uterine endometria. There was no significant difference in the levels between uterine endometrial cancers of G2 and G3, A and C, or stages II and III/IV and normal uterine endometria. Therefore, the active availability of PD-ECGF might contribute to the acceleration of angiogenic activity in the early process of invasion of well-differentiated uterine endometrial cancers.

Clinical significance of expression of estrogen receptor alpha and beta mRNAs in ovarian cancers.

Novel human estrogen receptor (ER)-β was identified in cDNA libraries from human testis. ER-β specifically expresses in testis, ovary, thymus, spleen, osteoblasts and fetus. ER-β might not conserve the same physiological functions as does ER-α. Therefore, the clinical significance of the expression of ER-α and ER-β mRNAs in ovarian cancers was investigated. The percentage of ER-β mRNA to ER-α mRNA ranged from 1.5 to 10% in normal ovaries. On the other hand, the ratios of ER-β mRNA to ER-α mRNA were in a wide range in ovarian cancers. There was no significant difference in the ratios among ovarian cancers classified according to histological types or clinical stages. In a 48-month survival rate, the patient prognosis in ovarian cancers with a low or high ratio of ER-β mRNA to ER-α mRNA (<1.5 or >10% of ER-β mRNA to ER-α mRNA) was significantly worse than that in ovarian cancers with a medium ratio (≧1.5 to ≦10% of ER-β mRNA to ER-α mRNA). In conclusion, the intact synchronized expression of ER-β mRNA interacting with ER-α mRNA might be damaged in some ovarian cancers, which might lead to poor patient prognosis.

Expression of estrogen receptor wild type and exon 5 splicing variant mRNAs in normal and endometriotic endometria during the menstrual cycle

Expression of estrogen receptor exon 5 splicing variant (ER-E5SV) mRNA contributes to dominant positive properties and metastatic potential of gynecological cancers. Therefore, in endometriotic endometria, which conserve implantation and tumor-like spreading potentials, the expression of ER-E5SV and estrogen receptor wild type (ER-WT) mRNA was analyzed, to investigate the biological implications. Estrogen receptor wild type mRNA in normal endometria was downregulated after ovulation, but not in endometriotic endometria. Therefore the wild type cascade in endometriotic endometria might be partly disorganized. The level of ER-E5SV mRNA in endometriotic endometria was lower than that in normal endometria, regardless of the stage in the menstrual cycle. However, there was no significant difference in the ratio of ER-E5SV to ER-WT mRNA in both normal and endometriotic endometria. On the other hand, the level of ER-E5SV mRNA in both normal and endometriotic endometria did not vary during the menstrual cycle. The dominant positive behavior of ER-E5SV might be masked by the functional cascade of ER-WT in normal endometria, but not in endometriotic endometria. This status might result in an incomplete response to endogenous steroids, and contribute to implantation and spreading potentials of endometriosis.