Osteopontin in Metastatic Breast Cancer Patients
Condition: Survival, Prosthesis
MeSH terms: Prosthesis Failure
Study type: Observational [Patient Registry]

Summary: 
      OPN plays an important role in various aspects of malignancy, particularly those involved in
      tissue invasion and metastasis, and OPN levels have been associated with aggressive¬ness in
      several cancer types, including breast cancer. Tumor response to treatment is a predictor of
      prognosis and overall survival for cancer patient population the investigators assigned
      osteopontin as potential contributors in breast cancer.
    

Detailed description: 
      Analysis of polymorphisms in the OPN regulatory region:

      The PCR amplification of the promoter regulatory region from the extracted DNA by using the
      following primers -3 (forward primer 5" -CAA GCT ACT GCA TAC TCG AAA TCA CA-3" ; reverse
      primer 5" - ACA ACC AAG CCC TCC CAG AAT TTA-3" ). PCR was performed using 50 ng DNA as a
      template under the following conditions: 95°C for 10 min, then 36 cycles of 94°C for 30 s, an
      annealing temperature for 60 s, and 72°C for 60 s, with a final extension at 72°C for 15 min.
      After affinity membrane purification using the QIAquick Gel Extraction kit (Qiagen, Carlsbad,
      CA, USA), the PCR products were subjected to cycle sequencing with the respective forward and
      reverse primer using an automated ABI 310 DNA sequencer.

      Measurement of serum level of osteopontin:

      OPN level were measured using Enzyme linked immnosorbent (ELISA) Kit supplied by IBL
      (Immuno-Biological laboratories Co. Ltd., Japan). 30 normal females were conducted with
      matched age with a median osteopontine serum level of 15 so, this level was assigned as a cut
      off value.

      Pretreatment serum level and polymorphism of oteopontine was correlated with different
      clinicopathological criteria of the patients, response to the treatment, PFS and OS.
    

Intervention name(s): Osteopontin
Intervention type(s): Diagnostic Test
Intervention description(s): Serum osteopontin (OPN) was measured by ELIZA and osteopontin gene mutation was analyzed by sequencing and correlated with clinicopathological criteria, response, PFS and OS.
Observational model: Cohort
Time perspective: Prospective
Sampling method: Probability Sample
Study population: 
        Denovo Metastatic breast
      
Biospecimen description: 
      The PCR amplification of the promoter regulatory region from the extracted DNA by using the
      following primers -3 (forward primer 5" -CAA GCT ACT GCA TAC TCG AAA TCA CA-3" ; reverse
      primer 5" - ACA ACC AAG CCC TCC CAG AAT TTA-3" ). PCR was performed using 50 ng DNA as a
      template under the following conditions: 95°C for 10 min, then 36 cycles of 94°C for 30 s, an
      annealing temperature for 60 s, and 72°C for 60 s, with a final extension at 72°C for 15 min.
      After affinity membrane purification using the QIAquick Gel Extraction kit (Qiagen, Carlsbad,
      CA, USA), the PCR products were subjected to cycle sequencing with the respective forward and
      reverse primer using an automated ABI 310 DNA sequencer.