Breast cancer fish lecture 1 Flashcards
HER 2 genes is also called the …………… (Erb-B2 receptor tyrosine kinase 2) gene
HER2 gene makes ………… proteins which are receptors on …………… cells
Normally, HER2 receptors help control cell gr……../di………/r……….
In about 2…% of breast cancers, the HER2 gene is amplified (makes too many copies of itself), therefore, does not work correctly
All these extra HER 2 genes tell breast cells to make …….. many HER 2 r…………….. (HER2 protein overexpression)
This makes breast cells grow and divide in an un………………. way
Breast cancers with HER2 genes amplification or HER2 protein overex…………. are called HER 2 p………..
HER2 -positive breast cancers tend to grow f………… and are more likely to meta………….. compared to HER2 negative breast cancers
But there are medicines specifically for HER 2 positive breast cancers
HER 2 genes is also called the ERBB2 (Erb-B2 receptor tyrosine kinase 2) gene
HER2 gene makes HER2 proteins which are receptors on breast cells
Normally, HER2 receptors help control cell growth/division/repair
In about 25% of breast cancers, the HER2 gene is amplified (makes too many copies of itself), therefore, does not work correctly
All these extra HER 2 genes tell breast cells to make too many HER 2 receptors (HER2 protein overexpression)
This makes breast cells grow and divide in an uncontrolled way
Breast cancers with HER2 genes amplification or HER2 protein overexpression are called HER 2 positive
HER2 -positive breast cancers tend to grow faster and are more likely to metastasise compared to HER2 negative breast cancers
But there are medicines specifically for HER 2 positive breast cancers
How we determine the status of HER2
- IHC testing (Immunohistochemistry finds out if there is ………. much HER2 ……….. in the cancer cells The results of the IHC test can be: 0 (negative). 1 + (also negative), 2_ (borderline), or 3 (…………. - HER2 protein overexpression)
- Fish Test (Fluoresence In situ Hybridization): finds out if there are too many ……….. of the HER2 gene in the cancer cells. The results of the FISH test can be positive (HER2 gene amp………….) or negative (no HER2 gene amplification)
sPoT0Light HER3 CISH test
(subtaction PRobe Technology Chromogenic In Situ Hybridization): finds out if there are too many …………. of the HER2 gene in the cancer cells. The results of the SPoT light test can be positive (HER2 gene amplification ) or negative (no HER2 gene amplification)
Inform HER2 Dual ISH test (Inform DUal In situ Hybridization): The inform HER2 Dual ISH test finds out if there are …………… many copies of the HER2 gene in the cancer cells. The results of the Inform HER2 Dual ISh test can be positive (HER2 gene amplification) or negative (no HER2 gene amplification)
How we determine the status of HER2
- IHC testing (Immunohistochemistry finds out if there is too much HER2 protein in the cancer cells The results of the IHC test can be: 0 (negative). 1 + (also negative), 2_ (borderline), or 3 (positive - HER2 protein overexpression)
- Fish Test (Fluoresence In situ Hybridization): finds out if there are too many copies of the HER2 gene in the cancer cells. The results of the FISH test can be positive (HER2 gene amplification) or negative (no HER2 gene amplification)
sPoT0Light HER3 CISH test
(subtaction PRobe Technology Chromogenic In Situ Hybridization): finds out if there are too many copies of the HER2 gene in the cancer cells. The results of the SPoT light test can be positive (HER2 gene amplification ) or negative (no HER2 gene amplification)
Inform HER2 Dual ISH test (Inform DUal In situ Hybridization): The inform HER2 Dual ISH test finds out if there are too many copies of the HER2 gene in the cancer cells. The results of the Inform HER2 Dual ISh test can be positive (HER2 gene amplification) or negative (no HER2 gene amplification)
HER2 is overexpressed in ..5% of cancer cells
HER2 is overexpressed in 25% of cancer cells
Why to test for HER2
Patient tumour> IHC 2+ fish/cish > He…………. therapy > if ….+ straight too Her……….. therapy
Patient tumour sample
Fish/CISH > + He……….. .therapy
Why to test for HER2
Patient tumour> IHC 2+ fish/cish > Herceptin therapy > if 3+ straight too Herceptin therapy
Patient tumour sample
Fish/CISH > + Herceptin therapy
Molecular subtypes of breast cancer
Based on the gens a cancer expresses:
- Lumical A breast cancer :ER+ and / or PR+. HER2, ……… levels of protein Ki-67; low-grade tend to grow …………. and have the best prognosis
Luminal B Breast cancer : ER+ and/or PR+ either HER2+/-. ……… levels of ki-67. Luminal B breast cancers generally grow slightly …… than luminal A cancer and their prognosis is slightly worse
Triple negative/basal like breast cancer ER, PR-, HER2; more common in women with B………. gene mutations
- HER2 enr……… breast cancer: ER,PR. HER2+ tend to grow fa………. than luminal cancers and can have ……….. prognosis, but they are often successfully treated with targeted therapies aimed at the HE…….. protein. such as Herceptin. Perjeta
(pertuzumab) , Tykerb (lapatinib, and Kadcyla (ado-trastuzumab emtansine) - normal like breast cancer:ER+ and/or PR+, …….. levels of the protein ki-67; has a g……. prognosis is slightly worse than Luminal A cancers prognosis, its prognosis, its prognosis is slightly worse than luminal A cancers prognosis
Molecular subtypes of breast cancer
Based on the gens a cancer expresses:
- Lumical A breast cancer :ER+ and / or PR+. HER2, low levels of protein Ki-67; low-grade tend to grow slowy and have the best prognosis
Luminal B Breast cancer : ER+ and/or PR+ either HER2+/-. high levels of ki-67. Luminal B breast cancers generally grow slightly faster than luminal A cancer and their prognosis is slightly worse
Triple negative/basal like breast cancer ER-,PR-, HER2; more common in women with BRCA1 gene mutations
- HER2 enriched breast cancer: ER,PR. HER2+ tend to grow faster than luminal cancers and can have worse prognosis, but they are often successfully treated with targeted therapies aimed at the HER2 protein. such as Herceptin (tratuzumab), Perjeta
(pertuzumab) , Tykerb (lapatinib, and Kadcyla (ado-trastuzumab emtansine) - normal like breast cancer:ER+ and/or PR+, low levels of the protein ki-67; has a good prognosis is slightly worse than Luminal A cancers prognosis, its prognosis, its prognosis is slightly worse than luminal A cancers prognosis
Luminal A breast cancer is hormone-receptor positive (estrogen-receptor and/or progesterone-receptor positive), HER2 negative, and has low levels of the protein Ki-67, which helps control how fast cancer cells grow. Luminal A cancers are low-grade, tend to grow slowly and have the best prognosis.
Luminal B breast cancer is hormone-receptor po……….(estro……..-receptor and/or proge………..-receptor positive), and either HER2 positive or HER2 negative with ……… levels of Ki-67. Luminal B cancers generally grow slightly faster than luminal A cancers and their prognosis is slightly worse.
Triple-negative/basal-like breast cancer is hormone-receptor negative (oe…………..-receptor and prog…………-receptor negative) and HER2 negative. This type of cancer is more common in women with BR…….. gene mutations. Researchers aren’t sure why, but this type of cancer also is more common among younger and African-American women.
HER2-enriched breast cancer is hormone-receptor ne………. (estrogen-receptor and progesterone-receptor negative) and HER2 pos…….HER2-enriched cancers tend to grow ………… than luminal cancers and can have a worse prognosis, but they are often successfully treated with targeted therapies aimed at the HER2 protein, such as Herceptin (chemical name: trastuzumab), Perjeta (chemical name: pertuzumab), Tykerb (chemical name: lapatinib), and Kadcyla (chemical name: T-DM1 or ado-trastuzumab emtansine).
Normal-like breast cancer is similar to luminal ….. disease: hormone-receptor positive (estrogen-receptor and/or progesterone-receptor positive), HER2 …………., and has l……… levels of the protein Ki-67, which helps control how fast cancer cells grow. Still, while normal-like breast cancer has a good prognosis, its prognosis is slightly worse than luminal A cancer’s prognosis.
Luminal A breast cancer is hormone-receptor positive (estrogen-receptor and/or progesterone-receptor positive), HER2 negative, and has low levels of the protein Ki-67, which helps control how fast cancer cells grow. Luminal A cancers are low-grade, tend to grow slowly and have the best prognosis.
Luminal B breast cancer is hormone-receptor positive (estrogen-receptor and/or progesterone-receptor positive), and either HER2 positive or HER2 negative with high levels of Ki-67. Luminal B cancers generally grow slightly faster than luminal A cancers and their prognosis is slightly worse.
Triple-negative/basal-like breast cancer is hormone-receptor negative (estrogen-receptor and progesterone-receptor negative) and HER2 negative. This type of cancer is more common in women with BRCA1 gene mutations. Researchers aren’t sure why, but this type of cancer also is more common among younger and African-American women.
HER2-enriched breast cancer is hormone-receptor negative (estrogen-receptor and progesterone-receptor negative) and HER2 positive. HER2-enriched cancers tend to grow faster than luminal cancers and can have a worse prognosis, but they are often successfully treated with targeted therapies aimed at the HER2 protein, such as Herceptin (chemical name: trastuzumab), Perjeta (chemical name: pertuzumab), Tykerb (chemical name: lapatinib), and Kadcyla (chemical name: T-DM1 or ado-trastuzumab emtansine).
Normal-like breast cancer is similar to luminal A disease: hormone-receptor positive (estrogen-receptor and/or progesterone-receptor positive), HER2 negative, and has low levels of the protein Ki-67, which helps control how fast cancer cells grow. Still, while normal-like breast cancer has a good prognosis, its prognosis is slightly worse than luminal A cancer’s prognosis.
SO… Main molecular alterations
Molecular genetics: amplification of ERBB2 (HER2, NEU) and mutations in BRCA1 and BRCA2
RAS proteins overexpression. TP… mutations RB… under expression, CC….. amplification
SO… Main molecular alterations
Molecular genetics: amplification of ERBB2 (HER2, NEU) and mutations in BRCA1 and BRCA2
RAS proteins overexpression. TP53 mutations RB1 under expression, CCND1 amplification
Fluorescene in SITU hybridisation (FISH) technique
Principles of technique
Steps that need to be considered
- Slide preparation: paraffin or frozen
- Pre-hybridisation steps: u,,,,,,,,,,,,,,,, target D,,,,,,,,,,
- Hybridisation of labelled pr,,,,,,,,,,,,,
- Post- hybridisation washes to remove non-hy,,,,,,,,,,,,, probes
- detection of pr,,,,,,,,,,
- Inter,,,,,,,,,,,,,
Fluorescene in SITU hybridisation (FISH) technique
Principles of technique
Steps that need to be considered
- Slide preparation: paraffin or frozen
- Pre-hybridisation steps: unmasking target DNA
- Hybridisation of labelled probes
- Post- hybridisation washes to remove non-hybridised probes
- detection of probe
- Interpretation
Paraffin embedded tissue pre-treatment and forzen tumour tissue pre-treatment
Fish protocol
1) Mark hybridizing a,,,,,,,,,
2) Apply f,,,,,,,, probe
3) Cover with c………. glass and seat with rubber …………
4) De…………
Hybridization
- incu…….
wash >
mounting
1) Apply D……… containing mounting me……….
2 ) Apply a g…….. cover
> imagine analysis
Paraffin embedded tissue pre-treatment and forzen tumour tissue pre-treatment
Fish protocol
1) Mark hybridizing area
2) Apply fish probe
3) Cover with cover glass and seat with rubber cement
4) Denature
Hybridization
- incubation
wash >
mounting
1) Apply DAPI containing mounting medium
2 ) Apply a glass cover
> imagine analysis
Metaphase spread pre-treatment
FISH protocol
1) Sample de………..
2) Dehy……….
3) FISH probe den………
4) Apply FISH probe on sample sl……….
5 ) Cover with cover gla……. and seal with rub……… .cameras
Hybridization
Hybridization
- incub……
wash >
mounting
1) Apply DAPI contai……. mo………. med………
2 ) Apply a glass c…….
> imagine an………3.
Metaphase spread pre-treatment
FISH protocol 1) Sample denature 2) Dehydrate 3) FISH probe denature 4) Apply FISH probe on sample slide 5 ) Cover with cover glass and seal with rubber cameras
Hybridization
Hybridization
- incubation
wash >
mounting
1) Apply DAPI containing mounting medium
2 ) Apply a glass cover
> imagine analysis
