Cancer platinum resistance detection and sensitization method
The phosphorylation status of the BAD protein is a determinant of ovarian cancer cell responsiveness to platinum chemotherapy. Indirect manipulation of BAD phosphorylation status influences cisplatin sensitivity. BAD phosphorylation represents a biomarker that predicts platinum sensitivity and is a therapeutic target to increase platinum sensitivity. The methods employ phospho-specific antibody against a particular amino acid residue or site. Phospho-specific protein characterization methods include immunohistochemical (IHC), flow cytometric, immunofluorescent, capture-and-detection, or reversed phase assay.
1. A method for determining cancer treatment sensitivity, comprising:
providing a cellular sample from an ovary of an ovarian cancer patient prior to platinum-based chemotherapy treatment;
forming a plurality of cell cultures from the cellular sample;
contacting the cell cultures with a plurality of predetermined amounts of the platinum-based chemotherapy treatment at more than one timepoint;
evaluating the phosphorylation status of BCL-2 Associated Death Promoter in the cell cultures, comprising:
isolating BCL-2 Associated Death protein from the cell cultures;
contacting the BCL-2 Associated Death protein from the cell cultures with at least one antibody, wherein the at least one antibody is anti-phospho-BAD ser 155; and
comparing the phosphorylation status of the ser155 amino acid residue on BCL-2 Associated Death Promoter in the cell cultures to a phosphorylation status of the ser155 amino acid residue on BCL-2 Associated Death Promoter on a platinum-based chemotherapy sensitive sample control;
wherein an elevated phosphorylation status of the BCL-2 Associated Death Promoter is indicative of platinum-based chemotherapy resistivity.
2. The method of claim 1 , wherein the cancer treatment is cisplatin.
3. The method of claim 1 , wherein the cellular samples are further collected by biopsies, ductal lavages, or fine needle aspirants.
4. The method of claim 1 , wherein the phosphorylation status of BCL-2 Associated Death Promoter is evaluated by a testing method selected from the group consisting of immunohistochemical, immunofluorescent, reverse-phase array, flow cytometric analysis, and tissue microarray.
5. The method of claim 4 , wherein the immunohistochemical testing method is performed comprising the steps
fixing and embedding cells from the cellular sample;
cutting sections from the embedded cells; and
staining the sections with antibodies against the at least one amino acid residue on BCL-2 Associated Death Promoter.
6. The method of claim 1 , further comprising evaluating the RNA expression status of PP2C, wherein the RNA expression is evaluated by
contacting protein, RNA, or cDNA from the cellular sample prior to platinum-based chemotherapy treatment with an antibody directed to PP2C; and
comparing the expression levels of the PP2C in the cellular sample to an expression levels of PP2C on a platinum-based chemotherapy-sensitive sample control;
wherein an elevated expression level of PP2C is indicative of platinum-based chemotherapy resistivity.
7. A method for determining cisplatin treatment sensitivity, comprising:
providing a cellular sample from an ovary of an ovarian cancer patient prior to cisplatin chemotherapy treatment;
forming a plurality of cell cultures from the cellular sample;
contacting the cell cultures with a plurality of predetermined amounts of the platinum-based chemotherapy treatment at more than one timepoint;
evaluating the phosphorylation status of BCL-2 Associated Death Promoter in the cell cultures using a testing method selected from the group consisting of immunohistochemical, immunofluorescent, reverse-phase array, flow cytometric analysis, and tissue microarray;
wherein the evaluating is performed by the steps comprising
isolating BCL-2 Associated Death protein from the cell cultures;
contacting the BCL-2 Associated Death protein with at least one antibody, wherein the at least one antibody is anti-phospho-BAD155; and
comparing the phosphorylation status of the ser155 amino acid residue on BCL-2 Associated Death Promoter in the cell cultures to a phosphorylation status of the ser155 amino acid residue on BCL-2 Associated Death Promoter on a cisplatin-sensitive sample control;
wherein an elevated phosphorylation status of the BCL-2 Associated Death Promoter is indicative of cisplatin resistivity.
8. The method of claim 7 , wherein the cellular samples are further collected by biopsies, ductal lavages, or fine needle aspirants.
9. The method of claim 7 , further comprising evaluating the RNA expression status of PP2C, wherein the RNA expression is evaluated by
contacting protein, RNA, or cDNA from the cellular sample prior to platinum-based chemotherapy treatment with an antibody directed to PP2C; and
comparing the expression levels of the PP2C in the cellular sample to an expression levels of PP2C on a cisplatin-sensitive sample control;
wherein an elevated expression level of PP2C is indicative of cisplatin chemotherapy resistivity.