Abstract
Purpose. Although pre-operative chemotherapy has improved the prognosis for individuals with osteosarcoma, approximately 40% of patients will die of their disease.The aim of this study was to quantitate proliferative activity in high grade osteosarcomas and to determine whether proliferation is a prognostic factor.
Patients. The study consisted of 27 patients with high grade non-metastatic osteosarcoma at various sites for whom pre-operative biopsies and resection specimens were available for review. All patients were treated similarly and had at least 24 months' follow-up from the date of diagnosis.
Methods. Proliferative activity (Ki-67 expression) was examined in the diagnostic biopsies immunohistochemically using the MIB-1 antibody. Proliferation was quantitated in two ways; (1) the number of immunopositive cells was counted manually using an ocular grid; or (2) the percentage of immunopositive nuclear area was assessed using morphometric image analysis. Proliferative index was evaluated in relation to patient outcome.
Results. Proliferative activity was seen in all biopsies.The median proliferative index as determined by counting cells was 24% (mean of 27%, range of 7–61%) and by image analysis was 2% (mean 3%, range 0.32–8.4).The correlation between MIB-1 proliferation indices determined either by image analysis methodology or manual cell counting was high (Spearman's rho=0.79). Proliferative index did not appear to predict either disease-free or overall survival.
Discussion. Tumor proliferation does not appear to be prognostic for high grade osteosarcomas.Whether assessment of this feature in conjunction with other tumor characteristics might be prognostic requires further study.
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- Aas T., Børresen A. L., Geisler S., Smith-Sørensen B., Johnsen H., Varhaug J. E., Akslen L. A., Lønning P. E. Specific P53 mutations are associated with de novo resistance to doxorubicin in breast cancer patients. Nat Med. 1996 Jul;2(7):811–814. doi: 10.1038/nm0796-811. [DOI] [PubMed] [Google Scholar]
- Baldini N., Scotlandi K., Barbanti-Bròdano G., Manara M. C., Maurici D., Bacci G., Bertoni F., Picci P., Sottili S., Campanacci M. Expression of P-glycoprotein in high-grade osteosarcomas in relation to clinical outcome. N Engl J Med. 1995 Nov 23;333(21):1380–1385. doi: 10.1056/NEJM199511233332103. [DOI] [PubMed] [Google Scholar]
- Benassi M. S., Molendini L., Gamberi G., Sollazzo M. R., Ragazzini P., Merli M., Magagnoli G., Sangiorgi L., Bacchini P., Bertoni F. Altered G1 phase regulation in osteosarcoma. Int J Cancer. 1997 Oct 21;74(5):518–522. doi: 10.1002/(sici)1097-0215(19971021)74:5<518::aid-ijc7>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
- Biesterfeld S., Klüppel D., Koch R., Schneider S., Steinhagen G., Mihalcea A. M., Schröder W. Rapid and prognostically valid quantification of immunohistochemical reactions by immunohistometry of the most positive tumour focus. A prospective follow-up study on breast cancer using antibodies against MIB-1, PCNA, ER, and PR. J Pathol. 1998 May;185(1):25–31. doi: 10.1002/(SICI)1096-9896(199805)185:1<25::AID-PATH40>3.0.CO;2-R. [DOI] [PubMed] [Google Scholar]
- Bubendorf L., Sauter G., Moch H., Schmid H. P., Gasser T. C., Jordan P., Mihatsch M. J. Ki67 labelling index: an independent predictor of progression in prostate cancer treated by radical prostatectomy. J Pathol. 1996 Apr;178(4):437–441. doi: 10.1002/(SICI)1096-9896(199604)178:4<437::AID-PATH484>3.0.CO;2-4. [DOI] [PubMed] [Google Scholar]
- Bubendorf L., Tapia C., Gasser T. C., Casella R., Grunder B., Moch H., Mihatsch M. J., Sauter G. Ki67 labeling index in core needle biopsies independently predicts tumor-specific survival in prostate cancer. Hum Pathol. 1998 Sep;29(9):949–954. doi: 10.1016/s0046-8177(98)90199-x. [DOI] [PubMed] [Google Scholar]
- Caulet S., Lesty C., Raphael M., Binet J. L., Diebold J. Quantitative study of KI-67 antibody staining in non-Hodgkin's lymphomas using image analysis. Anal Quant Cytol Histol. 1991 Aug;13(4):279–287. [PubMed] [Google Scholar]
- Chan H. S., Grogan T. M., Haddad G., Hipfner D. R., Deeley R. G., Cole S. P. Standardization of a single-cell assay for sensitive detection of multidrug resistance protein expression in normal and malignant cells in archival clinical samples. J Lab Clin Med. 1997 Sep;130(3):297–306. doi: 10.1016/s0022-2143(97)90025-1. [DOI] [PubMed] [Google Scholar]
- Choong P. F., Akerman M., Willén H., Andersson C., Gustafson P., Alvegård T., Rydholm A. Expression of proliferating cell nuclear antigen (PCNA) and Ki-67 in soft tissue sarcoma. Is prognostic significance histotype-specific? APMIS. 1995 Nov;103(11):797–805. doi: 10.1111/j.1699-0463.1995.tb01437.x. [DOI] [PubMed] [Google Scholar]
- Davis A. M., Bell R. S., Goodwin P. J. Prognostic factors in osteosarcoma: a critical review. J Clin Oncol. 1994 Feb;12(2):423–431. doi: 10.1200/JCO.1994.12.2.423. [DOI] [PubMed] [Google Scholar]
- Dictor M., Ehinger M., Mertens F., Akervall J., Wennerberg J. Abnormal cell cycle regulation in malignancy. Am J Clin Pathol. 1999 Jul;112(1 Suppl 1):S40–S52. [PubMed] [Google Scholar]
- Eastman A. Activation of programmed cell death by anticancer agents: cisplatin as a model system. Cancer Cells. 1990 Aug-Sep;2(8-9):275–280. [PubMed] [Google Scholar]
- Fan S., el-Deiry W. S., Bae I., Freeman J., Jondle D., Bhatia K., Fornace A. J., Jr, Magrath I., Kohn K. W., O'Connor P. M. p53 gene mutations are associated with decreased sensitivity of human lymphoma cells to DNA damaging agents. Cancer Res. 1994 Nov 15;54(22):5824–5830. [PubMed] [Google Scholar]
- Gerdes J., Li L., Schlueter C., Duchrow M., Wohlenberg C., Gerlach C., Stahmer I., Kloth S., Brandt E., Flad H. D. Immunobiochemical and molecular biologic characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki-67. Am J Pathol. 1991 Apr;138(4):867–873. [PMC free article] [PubMed] [Google Scholar]
- Glasser D. B., Lane J. M., Huvos A. G., Marcove R. C., Rosen G. Survival, prognosis, and therapeutic response in osteogenic sarcoma. The Memorial Hospital experience. Cancer. 1992 Feb 1;69(3):698–708. doi: 10.1002/1097-0142(19920201)69:3<698::aid-cncr2820690317>3.0.co;2-g. [DOI] [PubMed] [Google Scholar]
- Hendricks J. B., Wilkinson E. J. Quality control considerations for Ki-67 detection and quantitation in paraffin-embedded tissue. J Cell Biochem Suppl. 1994;19:105–110. [PubMed] [Google Scholar]
- Heslin M. J., Cordon-Cardo C., Lewis J. J., Woodruff J. M., Brennan M. F. Ki-67 detected by MIB-1 predicts distant metastasis and tumor mortality in primary, high grade extremity soft tissue sarcoma. Cancer. 1998 Aug 1;83(3):490–497. doi: 10.1002/(sici)1097-0142(19980801)83:3<490::aid-cncr18>3.0.co;2-r. [DOI] [PubMed] [Google Scholar]
- Hickman J. A. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev. 1992 Sep;11(2):121–139. doi: 10.1007/BF00048059. [DOI] [PubMed] [Google Scholar]
- Hitchcock C. L. Ki-67 staining as a means to simplify analysis of tumor cell proliferation. Am J Clin Pathol. 1991 Oct;96(4):444–446. doi: 10.1093/ajcp/96.4.444. [DOI] [PubMed] [Google Scholar]
- Irazusta S. P., Vassallo J., Magna L. A., Metze K., Trevisan M. The value of PCNA and AgNOR staining in endoscopic biopsies of gastric mucosa. Pathol Res Pract. 1998;194(1):33–39. doi: 10.1016/s0344-0338(98)80009-5. [DOI] [PubMed] [Google Scholar]
- Jensen V., Sørensen F. B., Bentzen S. M., Ladekarl M., Nielsen O. S., Keller J., Jensen O. M. Proliferative activity (MIB-1 index) is an independent prognostic parameter in patients with high-grade soft tissue sarcomas of subtypes other than malignant fibrous histiocytomas: a retrospective immunohistological study including 216 soft tissue sarcomas. Histopathology. 1998 Jun;32(6):536–546. [PubMed] [Google Scholar]
- King K. L., Cidlowski J. A. Cell cycle and apoptosis: common pathways to life and death. J Cell Biochem. 1995 Jun;58(2):175–180. doi: 10.1002/jcb.240580206. [DOI] [PubMed] [Google Scholar]
- Kirkegaard L. J., DeRose P. B., Yao B., Cohen C. Image cytometric measurement of nuclear proliferation markers (MIB-1, PCNA) in astrocytomas. Prognostic significance. Am J Clin Pathol. 1998 Jan;109(1):69–74. doi: 10.1093/ajcp/109.1.69. [DOI] [PubMed] [Google Scholar]
- Layfield L. J., Saria E. A., Berchuck A., Dodge R. K., Thompson J. K., Conlon D. H., Kerns B. J. Prognostic value of MIB-1 in advanced ovarian carcinoma as determined using automated immunohistochemistry and quantitative image analysis. J Surg Oncol. 1997 Dec;66(4):230–237. doi: 10.1002/(sici)1096-9098(199712)66:4<230::aid-jso2>3.0.co;2-c. [DOI] [PubMed] [Google Scholar]
- Lidang Jensen M., Schumacher B., Myhre Jensen O., Steen Nielsen O., Keller J. Extraskeletal osteosarcomas: a clinicopathologic study of 25 cases. Am J Surg Pathol. 1998 May;22(5):588–594. doi: 10.1097/00000478-199805000-00010. [DOI] [PubMed] [Google Scholar]
- Linden M. D., el-Naggar A. K., Nathanson S. D., Jacobson G., Zarbo R. J. Lack of correlation between flow cytometric and immunohistologic proliferation measurements of tumors. Mod Pathol. 1996 Jun;9(6):682–689. [PubMed] [Google Scholar]
- Link M. P., Goorin A. M., Horowitz M., Meyer W. H., Belasco J., Baker A., Ayala A., Shuster J. Adjuvant chemotherapy of high-grade osteosarcoma of the extremity. Updated results of the Multi-Institutional Osteosarcoma Study. Clin Orthop Relat Res. 1991 Sep;(270):8–14. [PubMed] [Google Scholar]
- Lowe S. W., Bodis S., McClatchey A., Remington L., Ruley H. E., Fisher D. E., Housman D. E., Jacks T. p53 status and the efficacy of cancer therapy in vivo. Science. 1994 Nov 4;266(5186):807–810. doi: 10.1126/science.7973635. [DOI] [PubMed] [Google Scholar]
- Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957–967. doi: 10.1016/0092-8674(93)90719-7. [DOI] [PubMed] [Google Scholar]
- Meikrantz W., Schlegel R. Apoptosis and the cell cycle. J Cell Biochem. 1995 Jun;58(2):160–174. doi: 10.1002/jcb.240580205. [DOI] [PubMed] [Google Scholar]
- Minimo C., McCue P. A., Pindzola A., Brennan J., Bibbo M. Role of computed quantitation of immunohistochemical staining of Ki-67 antigen in diagnosing ampullary lesions. Anal Quant Cytol Histol. 1996 Oct;18(5):400–404. [PubMed] [Google Scholar]
- Mochen C., Giardini R., Costa A., Silvestrini R. MIB-1 and S-phase cell fraction predict survival in non-Hodgkin's lymphomas. Cell Prolif. 1997 Jan;30(1):37–47. doi: 10.1046/j.1365-2184.1997.00066.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Molendini L., Benassi M. S., Magagnoli G., Merli M., Sollazzo M. R., Ragazzini P., Gamberi G., Ferrari C., Balladelli A., Bacchini P. Prognostic significance of cyclin expression in human osteosarcoma. Int J Oncol. 1998 May;12(5):1007–1011. [PubMed] [Google Scholar]
- Nawa G., Ueda T., Mori S., Yoshikawa H., Fukuda H., Ishiguro S., Funai H., Uchida A. Prognostic significance of Ki67 (MIB1) proliferation index and p53 over-expression in chondrosarcomas. Int J Cancer. 1996 Apr 22;69(2):86–91. doi: 10.1002/(SICI)1097-0215(19960422)69:2<86::AID-IJC3>3.0.CO;2-R. [DOI] [PubMed] [Google Scholar]
- Ohno T., Tanaka T., Takeuchi S., Matsunaga T., Mori H. Nucleolar organizer regions in bone tumors. Clin Orthop Relat Res. 1991 Nov;(272):287–291. [PubMed] [Google Scholar]
- Park H. R., Park Y. K. Expression of p53 protein, PCNA, and Ki-67 in osteosarcomas of bone. J Korean Med Sci. 1995 Oct;10(5):360–367. doi: 10.3346/jkms.1995.10.5.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petrilli A. S., Gentil F. C., Epelman S., Lopes L. F., Bianchi A., Lopes A., Figueiredo M. T., Marques E., De Bellis N., Consentino E. Increased survival, limb preservation, and prognostic factors for osteosarcoma. Cancer. 1991 Aug 15;68(4):733–737. doi: 10.1002/1097-0142(19910815)68:4<733::aid-cncr2820680412>3.0.co;2-0. [DOI] [PubMed] [Google Scholar]
- Radig K., Häckel C., Herting J., Oda Y., Mittler U., Neumann W., Roessner A. Expression of P-glycoprotein in high grade osteosarcomas with special emphasis on chondroblastic subtype. Gen Diagn Pathol. 1997 Feb;142(3-4):139–145. [PubMed] [Google Scholar]
- Raymond A. K., Chawla S. P., Carrasco C. H., Ayala A. G., Fanning C. V., Grice B., Armen T., Plager C., Papadopoulos N. E., Edeiken J. Osteosarcoma chemotherapy effect: a prognostic factor. Semin Diagn Pathol. 1987 Aug;4(3):212–236. [PubMed] [Google Scholar]
- Risio M. Methodological aspects of using immunohistochemical cell proliferation biomarkers in colorectal carcinoma chemoprevention. J Cell Biochem Suppl. 1994;19:61–67. [PubMed] [Google Scholar]
- Rudolph P., Peters J., Lorenz D., Schmidt D., Parwaresch R. Correlation between mitotic and Ki-67 labeling indices in paraffin-embedded carcinoma specimens. Hum Pathol. 1998 Nov;29(11):1216–1222. doi: 10.1016/s0046-8177(98)90248-9. [DOI] [PubMed] [Google Scholar]
- Sahin A. A., Ro J. Y., el-Naggar A. K., Wilson P. L., Teague K., Blick M., Ayala A. G. Tumor proliferative fraction in solid malignant neoplasms. A comparative study of Ki-67 immunostaining and flow cytometric determinations. Am J Clin Pathol. 1991 Oct;96(4):512–519. doi: 10.1093/ajcp/96.4.512. [DOI] [PubMed] [Google Scholar]
- Sallinen P. K., Haapasalo H. K., Visakorpi T., Helén P. T., Rantala I. S., Isola J. J., Helin H. J. Prognostication of astrocytoma patient survival by Ki-67 (MIB-1), PCNA, and S-phase fraction using archival paraffin-embedded samples. J Pathol. 1994 Dec;174(4):275–282. doi: 10.1002/path.1711740407. [DOI] [PubMed] [Google Scholar]
- Schmitt C. A., Lowe S. W. Apoptosis and therapy. J Pathol. 1999 Jan;187(1):127–137. doi: 10.1002/(SICI)1096-9896(199901)187:1<127::AID-PATH251>3.0.CO;2-T. [DOI] [PubMed] [Google Scholar]
- Scotlandi K., Serra M., Manara M. C., Maurici D., Benini S., Nini G., Campanacci M., Baldini N. Clinical relevance of Ki-67 expression in bone tumors. Cancer. 1995 Feb 1;75(3):806–814. doi: 10.1002/1097-0142(19950201)75:3<806::aid-cncr2820750310>3.0.co;2-s. [DOI] [PubMed] [Google Scholar]
- Serra M., Maurici D., Scotlandi K., Barbanti-Brodano G., Manara M. C., Benini S., Picci P., Bertoni F., Bacci G., Sottili S. Relationship between P-glycoprotein expression and p53 status in high-grade osteosarcoma. Int J Oncol. 1999 Feb;14(2):301–307. doi: 10.3892/ijo.14.2.301. [DOI] [PubMed] [Google Scholar]
- Shivji K. K., Kenny M. K., Wood R. D. Proliferating cell nuclear antigen is required for DNA excision repair. Cell. 1992 Apr 17;69(2):367–374. doi: 10.1016/0092-8674(92)90416-a. [DOI] [PubMed] [Google Scholar]
- Soomro I. N., Whimster W. F. Growth fraction in lung tumours determined by Ki67 immunostaining and comparison with AgNOR scores. J Pathol. 1990 Nov;162(3):217–222. doi: 10.1002/path.1711620307. [DOI] [PubMed] [Google Scholar]
- Thor A. D., Liu S., Moore D. H., 2nd, Edgerton S. M. Comparison of mitotic index, in vitro bromodeoxyuridine labeling, and MIB-1 assays to quantitate proliferation in breast cancer. J Clin Oncol. 1999 Feb;17(2):470–477. doi: 10.1200/JCO.1999.17.2.470. [DOI] [PubMed] [Google Scholar]
- Valente G., Giusti U., Kerim S., Gabriele P., Motta M., Ragona R., Navone R., Palestro G. High prognostic impact of growth fraction parameters in advanced stage laryngeal squamous cell carcinoma. Oncol Rep. 1999 Mar-Apr;6(2):289–293. doi: 10.3892/or.6.2.289. [DOI] [PubMed] [Google Scholar]
- Viberti L., Papotti M., Abbona G. C., Celano A., Filosso P. L., Bussolati G. Value of Ki-67 immunostaining in preoperative biopsies of carcinomas of the lung. Hum Pathol. 1997 Feb;28(2):189–192. doi: 10.1016/s0046-8177(97)90105-2. [DOI] [PubMed] [Google Scholar]
- Woosley J. T. Measuring cell proliferation. Arch Pathol Lab Med. 1991 Jun;115(6):555–557. [PubMed] [Google Scholar]
- van Oijen M. G., Medema R. H., Slootweg P. J., Rijksen G. Positivity of the proliferation marker Ki-67 in noncycling cells. Am J Clin Pathol. 1998 Jul;110(1):24–31. doi: 10.1093/ajcp/110.1.24. [DOI] [PubMed] [Google Scholar]