Targeting Cancer Stem Cells in Recurrent Lung Cancer
Grade Level at Time of Presentation
Senior
Major
Biology
Minor
Spanish
Institution
University of Louisville
KY House District #
36
KY Senate District #
36
Faculty Advisor/ Mentor
Sham Kakar, PhD
Department
Physiology
Abstract
Lung cancer is the leading cause of cancer mortality in the world. Five-year survivability still remains dismal due to drug resistance. The development of drug resistance with cisplatin or platinum/taxane combination chemotherapies has led patients to suffer from recurrent lung cancer. These chemotherapy treatments target cancer cells but leave behind cancer stem cells. Cancer stem cells (CSCs) are stem-like tumor cells that have the potential to differentiate, self-renew, and proliferate. This allows the cancer to relapse even after initial elimination of the tumor. We hypothesize that Verrucarin J will inhibit cell proliferation in lung cancer cells and CSCs by inducing apoptosis, targeting Wnt1 signaling pathway and reducing the expression levels of key stem cell markers and genes. To determine the effect of Verrucarin J, a MTT Assay was conducted to measure the cell proliferation rate and cell viability in the treated cells. This will then be used to define the IC50 of Verrucarin J. An apoptosis assay was performed to see if the compound induces cell death. To examine the relative levels of key markers found in cancer stem cells, such as ALDH1, Notch1, Hey 1, CD 133, Oct 4, B-catenin, Wnt1, TCF-4, and LGR5, RNA were amplified from the treated cells followed by real-time PCR. To establish a possible mechanism at which Verrucarin J was targeting the lung cancer cells, a GSK3 inhibitor (CHIR99021) was used to treat the A549 cells to affect the Wnt pathway. Results showed that Verrucarin J reduced cell viability in A549 and H1793 cells, decreased gene expression of key CSCs markers in a dose dependent manner and induced apoptosis in treated cells. From the PCR results of GSK3 inhibited cells, there was an increased levels of the Wnt pathway genes. This lead to a potential application of Verrucarin J as a therapeutic agent.
Targeting Cancer Stem Cells in Recurrent Lung Cancer
Lung cancer is the leading cause of cancer mortality in the world. Five-year survivability still remains dismal due to drug resistance. The development of drug resistance with cisplatin or platinum/taxane combination chemotherapies has led patients to suffer from recurrent lung cancer. These chemotherapy treatments target cancer cells but leave behind cancer stem cells. Cancer stem cells (CSCs) are stem-like tumor cells that have the potential to differentiate, self-renew, and proliferate. This allows the cancer to relapse even after initial elimination of the tumor. We hypothesize that Verrucarin J will inhibit cell proliferation in lung cancer cells and CSCs by inducing apoptosis, targeting Wnt1 signaling pathway and reducing the expression levels of key stem cell markers and genes. To determine the effect of Verrucarin J, a MTT Assay was conducted to measure the cell proliferation rate and cell viability in the treated cells. This will then be used to define the IC50 of Verrucarin J. An apoptosis assay was performed to see if the compound induces cell death. To examine the relative levels of key markers found in cancer stem cells, such as ALDH1, Notch1, Hey 1, CD 133, Oct 4, B-catenin, Wnt1, TCF-4, and LGR5, RNA were amplified from the treated cells followed by real-time PCR. To establish a possible mechanism at which Verrucarin J was targeting the lung cancer cells, a GSK3 inhibitor (CHIR99021) was used to treat the A549 cells to affect the Wnt pathway. Results showed that Verrucarin J reduced cell viability in A549 and H1793 cells, decreased gene expression of key CSCs markers in a dose dependent manner and induced apoptosis in treated cells. From the PCR results of GSK3 inhibited cells, there was an increased levels of the Wnt pathway genes. This lead to a potential application of Verrucarin J as a therapeutic agent.