J Korean Surg Soc.  2011 Jun;80(6):404-411. 10.4174/jkss.2011.80.6.404.

Expression of the survivin-2B splice variant related to the progression of colorectal carcinoma

• Cho GS
• Ahn TS
• Jeong D
• Kim JJ
• Kim CJ
• Cho HD
• Park DK
• Baek MJ

• KMID: 2212185
• DOI: http://doi.org/10.4174/jkss.2011.80.6.404

Abstract

PURPOSE
Recently, two alternatively spliced survivin variants, survivin-DeltaEx3 and survivin-2B, were identified in a single copy of the survivin gene. It has been reported that the expressions of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma. We investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and analyzed correlations between survivin expression levels and clinicopathologic features.
METHODS
We used Western blot and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to analyze the protein and mRNA expression levels of survivin variants in 51 colorectal carcinomas. The quantitative RT-PCR was performed using primer pairs specific for survivin and each of its splice variants, then normalized for the gene that encodes glyceraldehydes-3-phosphate dehydrogenase.
RESULTS
In Western blotting, the protein levels of survivin were higher in the tumor tissue than in normal tissue. The expression of survivin, survivin-2B and survivin-DeltaEx3 mRNA was present in 96%, 64.7%, and 82.4% of the samples, respectively. When the pathologic parameters were compared, colorectal cancers of advanced pT stages showed significant decrease in survivin-2B mRNA expression by the quantitative RT-PCR (P < 0.001).
CONCLUSION
The decreased expression of survivin-2B might be related to tumor progression in colorectal cancers. This finding indicates that alternatively spliced variants of survivin may be involved in refining the functions of survivin during tumor progression.

Keyword

Survivin variant; Colorectal neoplasms; Quantitative RT-PCR

MeSH Terms

Blotting, Western
Coat Protein Complex I
Colorectal Neoplasms
Humans
Polymerase Chain Reaction
Reverse Transcription
RNA, Messenger
Coat Protein Complex I
RNA, Messenger

Figure

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  Fig. 1 Western blotting for survivin in colon cancer tissues. Normal colon (N) and colon cancer (T) tissues were analyzed in a 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis gel. Single and distinct bands of approximately 18.1 kDa are evident, indicating the presence of the survivin protein.

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  Fig. 2 Specificities of the amplification products detected by agarose gel electrophoresis, with distinct bands of the calculated sizes for all polymerase chain reaction (PCR) products at the end-point of 50 PCR cycles. GAPDH, glyceraldehydes-3-phosphate dehydrogenase.

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  Fig. 3 GAPDH-normalized mRNA levels of survivin and its variants in colorectal cancers in relation to the different variables. A significant decrease in the expression of survivin-2B is noted in advanced T stage (pT3) patients. Top line representslargest value, bottom line represents smallest value, the upper line of the box represents upper quartile, the lower line of the box represents lower quartile, and inner line of the box represents median value.

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  Fig. 4 Ratios of the relative mRNA levels between survivin and its variants in colorectal cancer. No significant change is evident in relation to T stage. Top line represents largest value, bottom line represents smallest value, the upper line of the box represents upper quartile, the lower line of the box represents lower quartile, and inner line of the box represents median value.

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