Field of the Invention:

This invention relates to the method of inducing and stimulating pancreatic cancer stem cells to normal cancer cells and apoptosis of both pancreatic cancer stem cells and cancer cells which can be used in institutions such as medicine, molecular biology, a pharmaceutical industry where cancer treatment studies and treatments are performed.

State of the Art: Prevention of the spread of cancer, which is one of the most important health problems of our age, and of pancreatic cancer, which is one of the leading aggressive tumors, is one of the important health studies. The pancreas, which has many important functions in the body, has two main functions: digestion of food and regulation of blood sugar. Therefore, the pancreas has an important role in the health balance of the body. Pancreatic Cancer is grouped as initial (Carcinoma in situ), Stage I, Stage II, Stage III and Stage IV. This classification is made according to tumor size, lymph node involvement, and then spread to other parts / organs of the body. Surgery, radiation therapy, chemotherapy, cherno-radio therapy and targeted therapies are used against pancreatic cancer

In the treatment of pancreatic cancer, treatment options are created by considering the stage of the tumor. Chemotherapeutics frequently used in treatment are active substances such as Paclitaxel, 5-Fluorouracil, Gemcitabine Hydrochloride, Mitomycin C, Erlotinib Hydrochloride, Irinotecan Hydrochloride. Despite the current treatment options, pancreatic cancer treatment is not very successful and pancreatic cancer is still called the "Silent Killer". Stem cells, on the other hand, are cells found in the bodies of higher eukaryotes such as humans and animals that can transform into other cell types depending on the need. Cells in organs and tissues are formed by the differentiation of stem cells, and in case these cells are damaged or need renewal, they are renewed by the differentiation of stem cells. In other words, body cells in the body called somatic cells are formed by the differentiation of stem cells.

Although the concept of stem cells indicates a positive situation in general, this concept indicates a completely negative situation in formations such as cancer. The cells in the tumor mass are damaged due to their intense metabolic activities or due to the chemotherapeutics applied. The continuity of these tumor cells is provided by 'cancer stem cells' in the tumor mass, which are stem cells. The ability of these stem cells to be directed to cell death mechanisms such as apoptosis or to differentiate into tumor cells is one of the important issues in cancer molecular biology, and there is not much information about the induction of these cells to the said apoptotic pathway or cell differentiation. While induction of tumor cells to apoptosis is one of the most important principles in drug development studies, inhibition studies for these cells with stem cell characteristics after "cancer stem cell" modeling have become a very important need.

Application US8491896B2 describes the effect of anti-use compositions and methods of use on pancreatic cancer. The antibodies in question show some novel and beneficial therapeutic properties, such as binding with high specificity.

Application AU2012290949B2 describes a pharmaceutical composition for the treatment and/or prevention of pancreatic cancer. New use of an antibody against CAPRIN-1 in pancreatic cancer or a drug fragment thereof, such as a therapeutic and/or preventive agent.

Studies aimed at revealing and inhibiting the metabolic functions of cancer stem cells become even more important after the development of powerful chemotherapeutics. While normal tumor cells are eliminated with chemo or radiotherapy, stem cells remain resistant and these cells show a more aggressive course. In the applications mentioned above, they fall short of treating pancreatic cancer. In the treatment of pancreatic cancer patients, stopping the course of the disease, initiating cell death in cancerous cells and cancerous stem cells, initiating the transformation of cancerous stem cells into healthy cells is not possible with current treatment methods. There is a need for a method that prevents pain and side effects in the treatment of pancreatic patients.

Description of the Invention:

In the treatment of pancreatic cancer patients, there is a need for a method that stops the course of the disease, initiates cell death in cancer cells and cancer stem cells, and prevents pain and side effects in the treatment of pancreatic patients. Fentanyl is an effective substance used to stop pain in cancer.

The present invention is the effect of fentanyl on inducing apoptosis, a mechanism of cell death in pancreatic tumor cells PANC-1 and BxPC-3 cells, and inducing cancer stem cells into normal cancer cells or apoptosis.

Disclosure of the Invention:

In the invention, the active ingredient fentanyl induces apoptosis in pancreatic tumor cells PANC-1 and BxPC-3. However, it also stimulates cancer stem cells in the tumor cell population to both apoptosis and non-stem cell tumor cells.

According to the result obtained by flow cytometry analysis, it is characterized by the fact that while the amount of cancer stem cells in PANC-1 cell was 3.2%, it decreased to 1.5% after the application of 5-100 micromole (pmol) fentanyl. It is characterized by the fact that the amount of cancer stem cells in BxPC-3 cell decreased to 1.3% after the application of 5-100 pmol fentanyl, while the amount of cancer stem cells was 2.2% according to the result obtained by flow cytometry analysis. It has been shown by Real Time PCR method and Western Blot analysis that both mRNAs and protein expressions of the stem cell markers Sox2, Nanog and Oct4 genes are reduced after 5-100 micromole (pmol) application in PANC-1 and BxPC-3 cell populations.

Detailed Disclosure of the Invention:

Pancreatic cancer comes first among aggressive tumors. Pancreatic Cancer is grouped as initial (Carcinoma in situ), Stage I, Stage II, Stage III and Stage IV. This classification is made according to tumor size, lymph node involvement, and then spread to other parts/organs of the body. Surgery, radiation therapy, chemotherapy, chemo-radio therapy and targeted therapies are used against pancreatic cancer.

In the treatment of pancreatic cancer, treatment options are created by considering the stage of the tumor. Chemotherapeutics commonly used in treatment; Paclitaxel, 5- Fluorouracil, Gemcitabine Hydrochloride, Mitomycin C, Erlotinib Hydrochloride, Irinotecan Hydrochloride. Despite the current treatment options, pancreatic cancer treatment is not very successful and pancreatic cancer is still called the "Silent Killer". In the invention, the active ingredient fentanyl induces apoptosis in pancreatic tumor cells PANC-1 and BxPC-3. However, it also stimulates cancer stem cells in the tumor cell population to both apoptosis and non-stem cell tumor cells.

According to the result obtained by performing flow cytometry analysis, it is characterized by the fact that while the amount of cancer stem cell in PANC-1 cell is 3.2%, it has decreased to 1.5% after 5-100 pmol fentanyl application. It is characterized by the fact that the amount of cancer stem cells in BxPC-3 cell decreased to 1.3% after the application of 5-100 pmol fentanyl, while the amount of cancer stem cells was 2.2% according to the result obtained by flow cytometry analysis.

In the mRNA expression studies by RealTime PCR, it is revealed that Fentanyl application increases the mRNA expression of genes favoring apoptosis (such as Bax, Bad) and causes a decrease in mRNAs of genes against apoptosis (Bcl-2). Similarly, it is shown by Western Blot method that the expression levels of proteins that play a role in favor of apoptosis increase and that the expression levels of proteins against apoptosis decrease.

It is shown by RealTime PCR and Western Blot methods that the expression of the stem cell markers Sox2, OCT4 and Nanog genes decreased both at the mRNA level and at the protein level after fentanyl administration.