Anne Zielinski  

Home Institution
Albert-Ludwigs-Universität Freiburg

Host Institution
California Pacific Medical Center Research Institute (CPMC-RI)
Research Mentors: Pierre-Yves Desprez, PhD; Sean Mc Allister, PhD

E-Mail: AnneZielinski@gmx.de
Research Topic
see Abstract
Personal Reactions to the U.S. Experience
If you are going to San Francisco you can be sure that you are going to live in one of the most beautiful and charming cities in the US! There are so many things to do, that you don’t know what to do first. I have never experienced time running by so fast. Everything and everyone seems to move faster… well, except the public transportation, which is pretty good for an American city, but still moves very slowly.
I chose to live in Berkeley and work in San Francisco, which has the advantage that you are surrounded by students, live in a smaller city with many bars, and are very close to nature
During my time here, I have enjoyed going on tremendous bike rides in the Bay Area. Before I came here, I never thought, that I would ever ride a bike in the US, but yes, we do have bike lanes here and many people incorporate bike riding into their daily lifestyle. Sport plays a very important role here, most people try to live healthily, by being involved in a lot of sports (and ironically driving their cars to go most everywhere) and eating organic produce.
This is only one thing that shows that San Francisco is a very unique place in the US, but almost everything is special here. The Bay Area contains a huge variety of people not only regarding their ethnic backgrounds, but also their political points of view and their lifestyles. The area is very liberal in spirit and everybody freely expresses their opinion here. Generally, if there is a protest about something in the US, it is probably going to start here.
I have enjoyed working in the lab very much. I haven’t ever felt so comfortable at a place from the very first second. Everybody is extremely friendly and is willing to help you with whatever you need. Ironically, most of my coworkers were international, like myself, so in a sense I learned more about Americans outside of work. In fact, I missed American people, since they were in the minority at the lab, but I got to know many different cultural backgrounds, lifestyles, and accents!
Greatest Difficulties Encountered
Rescheduling my flight back to Germany with Delta: I booked my return flight to Germany with Delta a year in advance but since Delta hadn’t prepared flight schedules that far in advance, I chose a random date for my return flight with the intention of rescheduling the flight closer to the date of departure. But Delta wasn’t able to reschedule my paper ticket, so I had to go to the airport twice trying to resolve this problem! Conclusion: Avoid Delta if possible!
Most humorous incident
Once, the ice machine in the lab broke and we were supposed to schedule our experiments according to this inconvenience. Well, of course we didn’t want to do that, so we all went to Walgreens and bought cubed ice. You can imagine the gaze of the salesperson!
Helpful Hints for Future Students
- Choose your room through craigslist, but be very careful in the Bay Area. You can find very special housing and people that live a very special lifestyle.
- Get a Safeway card, otherwise groceries will be even more expensive than it is already in the Bay Area.
- Open a bank account at Deutsche Bank and at Bank of America. Get cash out of the ATM with your Deutsche Bank card for free and then make a deposit on your Bank of America account.
- Check out the outlet stores and get good deals for brands.
- Buy a Metro PCS cell phone plan including a month to month flat rate, and call Germany for $0.03 per minute. Disadvantage: The cell phones you have to buy have very bad batteries.
- Go to a Giant’s game at the AT&T stadium and enjoy the view of the Bay.

  table of contents

 

Abstract on Research Topic

Inhibition of cancer cell aggressiveness by cannabiniods
Authors: Anne Zielinski

Institution:
California Pacific Medical Center

Introduction:
Cannabis sativa consists of over 60 cannabinoids and ∆-9 tetrahydrocannabinol (THC) is the major psychoactive component. THC has been shown to be an inhibitor of many cancer types by inducing apoptosis, reducing cell proliferation and invasiveness. The effect of THC is mainly due to the activation of two G-protein coupled receptors, CB ₁ and CB ₂. However, the anticancer effect of additional cannabinoids has been proposed to the result of interaction independent of CB1 and CB2. The clinical use of THC is fairly limited, since the interaction with the CB₁ receptor results in a psychotropic effect. Therefore, we are looking for cannabinoids that inhibit cancer cell growth without activating CB ₁ receptors and consequently lack the psychotropic effect. Cannabidiol (CBD) exists in reasonable amounts in cannabis sativa and interacts at an insignificant level with CB ₁ and CB ₂.

Purpose and Methods:

Project 1:
Looking for the signaling pathway through which CBD mediates its antitumor effect, we came across the inhibitor of differentiation, Id-1. Id proteins belong to the family of basic- helix-loop-helix (bHLH) proteins and are inhibitors of differentiation by preventing the binding of other bHLH factors to the DNA. Increased expression of Id-1 has been shown to correlate with cancer cell aggressiveness.
In order to find a more potent drug than CBD and to figure out the main structural component of CBD that is responsible for the down regulation of Id-1, a collaborating group is synthesizing cannabinoids for us according to our suggestions.
We use human breast cancer cell lines and treat them for 3 days in RPMI and 0.1% FBS with each CBD analog. We analyze the Id-1 expression by Western Blot analysis and determine the inhibition of Id-1 on the transcription level by RT- PCR. To test the antiproliferative effect of the CBD analogs we use MTT assay. In order to evaluate the effect of CBD analogs on tumor invasion, we use Boyden Chamber Invasion assay.
Results: In progress

Project 2:
In order to find non psychotropic inhibitors of cancer cells, our group discovered that the addition of CBD to THC has synergistic effect in cancer cell growth. Therefore we treated several human Glioblastoma (GBM) cell lines with THC in combination with a four- fold lower concentration of CBD for 3 days in RPMI and 0.1% FBS. For cell cycle analysis we used FACS analysis using Cell Quest Pro and Modfit software. Other members of our group investigated proliferation using MTT assay, apoptosis using fluorescein-tagged UTP and PI by use of Apo-Direct apoptosis kit and Western Blot for analyzing the expression of ERK and caspases.
Results:
It was found, that CBD and THC act synergistically in reducing cell growth, upregulates significantly apoptosis in two out of three GBM cell lines but does not synergistically inhibit invasion of U251 GBM cells. We found, that the inhibitory effect of the combination treatment is due to activation of CB₂ receptors, inhibition of ERK and the G1/S transition in cell cycle.