PAI1 Regulates Cell Morphology and Migration Markers in Trastuzumab-Resistant HER2-Positive Breast Cancer Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture, Transfection, Drug Treatment
2.2. Combination Therapy Experiment
2.3. Quantitative Real-Time PCR
2.4. Gene Expression Heatmap
2.5. Western Blot
2.6. Immunofluorescence
2.7. Statistical Analysis
3. Results
3.1. TGF-β and Migration Responsive Genes Regulated by Acquired Resistance in HER2-Positive Trastuzumab Resistant Cell Lines
3.2. PAI1 Regulates Migration Responsive Gene Expressions in Parental and Resistant HER2-Positive Cell Lines
3.3. PAI1 Regulates Cell Morphology in Parental and Resistant HER2-Positive Cell Lines
3.4. Synergistic Effect of Trastuzumab+Aleplasinin on HER2-Positive Breast Cancer Cells
3.5. Trastzumab+Aleplasinin Combination Decrease PAI1 Expression
3.6. Trastzumab+Aleplasinin Combination Decrease Migration Responsive Marker Expressions
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Cell Lines | p-IC50 (μM) | R-IC50 (μM) |
---|---|---|
SKBR3 | 7.5 ± 0.6 | 6.4 ± 0.6 |
HCC1954 | 10.2 ± 0.8 | 12.5 ± 1.1 |
Gene Name | Sequence | Product Length |
---|---|---|
WWP1 | F: GCAGCTCATCTCCAACCATAG | 201 bp |
R: GAGACGGAGATGAAGGTGTG | ||
CARM1 | F: CAGTTTTATGGCTACCTGTCCC | 150 bp |
R: AAACGACAGGATCCCAGAGC | ||
RASGM1 | F: AAATCAATGCCCGTGACTGG | 166 bp |
R: TCAGAGAACGATATCCTCCGG | ||
THBS1 | F: GCTCTACCAGTGTCCTCCTC | 213 bp |
R: TCTCTTCAGTCACTTTGCGG | ||
KCDT1 | F: AGGAGTGTTGGAGGAAGCAG | 193 bp |
R: CAACTGCTCGAACTTCCAGC | ||
SGCA | F: CAGGTCATTGAGGTCACAGC | 183 bp |
R: CCCCAAGGCTGAGAGGAAG | ||
MCAM | F: ACTGGTTTTCTGTCCACAAGG | 203 bp |
R: GATGCGGTACTCCTGGGAC | ||
FXR2 | F: GGGGATGAAGTGGAGGTTTATTC | 177 bp |
R: AAGGGGATTGGGATTAACTGG | ||
MTMR3 | F: TTCCCAGGAAGCAGCTGATC | 249 bp |
R: ACTGACACCTGATAACTTTGCAG | ||
SOCS3 | F: GAGAGCGGCTTCTACTGGAG | 162 bp |
R: CTGGATGCGCAGGTTCTTG | ||
SLC2A4 | F: AGAGCCACCCCAGGAAAAG | 225 bp |
R: CGGAGAGGACTGGTCACTG | ||
MMP2 | F: CAGGATCATTGGCTACACACC | 151 bp |
R: CCAGCGGCCAAAGTTGATC | ||
MMP9 | F: TGGATCCAAAACTACTCGGAAG | 250 bp |
R: CATCGTCGAAATGGGCGTC | ||
PAI1 | F: AGACCGATTATTGACCGACC | 210 bp |
R: CCAGTTGTGAGATCCGCTAC | ||
TIMP1 | F: GACCACCTTATACCAGCGTTATG | 183 bp |
R: CCATCCTGCAGTTTTCCAGC | ||
TGIF1 | F: AGGTCTGTAACTGGTTCATCAAC | 231 bp |
R: GAGACAGTGGCCTCCCTAG | ||
COL6A1 | F: AAGTCTTCTCGGTGGCCATC | 198 bp |
R: CGAAGGAGCAGCACACTTG | ||
AAK1 | F: TGAGGGATCTTTCAGGGCAC | 208 bp |
R: GGCAACAGCTTCACAGGTATC | ||
MAN1A1 | F: ACAGTGGGGTCAACATCATG | 155 bp |
R: TGCGGATCAAATGAGTCTCG | ||
NT5C2 | F: TGCTGTGTACAAGTCCCCAG | 184 bp |
R: RGAGGTTTCCATAGGCATCGAC | ||
IRS1 | F: TCTTCTTCATCGAGGTGGGC | 170 bp |
R: CCCCGACGATTGGCTCTTAC | ||
KCNIP3 | F: GGGCTTTAAGAATGAGTGTCCC | 151 bp |
R: AAAGTGGATGGCCCCGTTC | ||
C4BPA | F: TCTACAAACGATGCAGACACC | 236 bp |
R: GCTGTGCCTTCCATTCCTG | ||
PSG1 | F: TCACCTTACACCTGGAGACTC | 159 bp |
R: AGCTGTGAGTCATAGGGAGG | ||
ALAD | F: AGAGTTCCCAAGGACGAGC | 172 bp |
R: ATGCTCCGTTTTCACTCAGG | ||
FGD1 | F: AGAAGTTTGAAAGAGAGCCTGTG | 214 bp |
R: CTATCAATGCCGCTGTCCCG | ||
TFAP2B | F: GTCCAGTCAGTTGAAGATGCC | 165 bp |
R: GACGGAGCAAAACACCTCG | ||
AHCYL1 | F: CGCTGGTCTGCTTGTAACATC | 175 bp |
R: CCCCATCATCCAGGATCATG | ||
HSPG2 | F: AGATGGTTTATTTCCGAGCCC | 181 bp |
R: AGCTCCTTGATGAACACCAC | ||
IL1RAP | F: CGACTATCACTTGGTATATGGGC | 164 bp |
R: CCTGGTGAGATGAAACGTACG | ||
EIF3S6 | F: GGACAAGCATGGTTTTAGGC | 186 bp |
R: TGCATTAAGATTTCAGAGGCCAG | ||
HSP47 | F: CGCCATGTTCTTCAAGCCAC | 244 bp |
R: CTTTTCAAGGCGCTCGAGAG | ||
MIDORI | F: ACAGCGACGTCAGGTTCAC | 169 bp |
R: GCATCTTCTTCTCGACACCTG | ||
GAPDH | F: ACAGTTGCCATGTAGACC | 152 bp |
R: TTGAGCACAGGGTACTTTA |
Cell Lines | Drug Combo CI Value | Description of Effect |
---|---|---|
SKBR3-P | 1.06 | Nearly additive |
SKBR3-R | 0.00 | Very strong synergism |
HCC1954-P | 0.92 | Nearly additive |
HCC1954-R | 0.49 | Synergism |
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Boz Er, A.B.; Er, I. PAI1 Regulates Cell Morphology and Migration Markers in Trastuzumab-Resistant HER2-Positive Breast Cancer Cells. Life 2024, 14, 1040. https://doi.org/10.3390/life14081040
Boz Er AB, Er I. PAI1 Regulates Cell Morphology and Migration Markers in Trastuzumab-Resistant HER2-Positive Breast Cancer Cells. Life. 2024; 14(8):1040. https://doi.org/10.3390/life14081040
Chicago/Turabian StyleBoz Er, Asiye Busra, and Idris Er. 2024. "PAI1 Regulates Cell Morphology and Migration Markers in Trastuzumab-Resistant HER2-Positive Breast Cancer Cells" Life 14, no. 8: 1040. https://doi.org/10.3390/life14081040