SiRNA targeting ETS1 and ELK1 and method of using same in the inhibition of CIP2A gene in cancer treatment
Disclosed are methods of attenuating activity of the gene promoter of CIP2A. siRNAs are used to target against Ets1 and Elk1 transcriptional factors, thereby blocking the binding of Ets1 and Elk1 to the CIP2A gene promoter. It is disclosed that the siRNAs targeted against Ets1 and Elk1 attenuate the gene expression of CIP2A. A kit containing siRNA reagents for attenuating the CIP2A gene expression is also disclosed.
1. A method of inhibiting gene expression of CIP2A in a cell, comprising the steps of:
(i) providing a cell selected from the group consisting of liver cancer cell, endometrial cancer cell and cervical cancer cell;
(ii) providing a first siRNA targeted against Ets1 mRNA, said first siRNA hybridizes to a target sequence of Ets1 mRNA selected from the group consisting of SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37;
(iii) providing a second siRNA targeted against Elk1 mRNA, said second siRNA hybridizes to a target sequence of Elk1 mRNA selected from the group consisting of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO: 41; and
(iv) exposing said first siRNA and said second siRNA to said cell, wherein said first siRNA and said second siRNA together inhibit the gene expression of CIP2A in said cell.
2. The method of claim 1 , wherein said first siRNA is at least one siRNA selected from the group consisting of SEQ ID NO: 30 and SEQ ID NO: 31.
3. The method of claim 1 , wherein said second siRNA is at least one siRNA selected from the group consisting of SEQ ID NO: 32 and SEQ ID NO: 33.
4. A method for inhibiting gene expression of CIP2A in a cell in vitro, said method comprising the steps of:
(a) introducing into a cell a first siRNA, said cell is selected from the group consisting of liver cancer cell, endometrial cancer cell and cervical cancer cell, wherein said first siRNA is at least one siRNA selected from the group consisting of SEQ ID NO: 30 and SEQ ID NO: 31;
(b) introducing into said cell a second siRNA, wherein said second siRNA is at least one siRNA selected from the group consisting of SEQ ID NO: 32 and SEQ ID NO: 33; and
(c) culturing said cell to inhibit gene expression of CIP2A, wherein said inhibition of gene expression of CIP2A is evidenced by a decrease in CIP2A mRNA transcript.
5. A method for inhibiting gene expression of CIP2A in a human in vivo, comprising the steps of:
(i) providing a first siRNA targeted against Ets1 mRNA, said first siRNA hybridizes to a target sequence of Ets1 mRNA selected from the group consisting of SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37;
(ii) providing a second siRNA targeted against Elk1 mRNA, said second siRNA hybridizes to a target sequence of Elk1 mRNA selected from the group consisting of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO: 41; and
(iii) administering to a human an effective amount of said first siRNA targeted Ets1 mRNA and said second siRNA targeted against Elk1 mRNA, said human suffers from a cancer selected from the group consisting of liver cancer, endometrial cancer and cervical cancer, wherein said first siRNA and said second siRNA together inhibit said gene expression of CIP2A in said human.
6. The method of claim 5 , wherein said first siRNA consists of a nucleotide sequence set forth in SEQ ID NO: 30 or SEQ ID NO: 31.
7. The method of claim 5 , wherein said second siRNA consists of a nucleotide sequence set forth in SEQ ID NO: 32 or SEQ ID NO: 33.
8. The method of claim 5 , wherein said human is suffering from cervical cancer or endometrial cancer.