Protein Kinase C-β Gene Variants, Pathway Activation, and Enzastaurin Activity in Lung Cancer

Sang-Haak Lee; Tingan Chen; Jun Zhou; Jennifer Hofmann; Gerold Bepler

Clin Lung Cancer. 2010;11(3)

Conclusion

Apparently PKCβ2 does not possess sequence alterations that lead to enzastaurin hypersensitivity. The phosphorylation inhibition of PKCβ2, GSK3β, S6RP, Akt, and FKHR may constitute a useful pharmacodynamic parameter for in vivo target inhibition by enzastaurin.

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Sang-Haak Lee and Tingan Chen contributed equally to this work.

Acknowledgments
This work was supported in part by grants U01-CA101222 and P50-CA119997 to G. B. from the National Cancer Institute.

Cite this: Protein Kinase C-β Gene Variants, Pathway Activation, and Enzastaurin Activity in Lung Cancer - Medscape - May 01, 2010.

Table 1. Primers Used for Sequencing of *PRKCB1* Gene in Lung Cancer
Exon	Forward Primer	Reverse Primer	Amplicon Size
1	GCGTGCAGAATGACCAATGGGATGGA	GGAGCCGGAGCCCGAGAGG	594
1	CAGCAGCTGGGCGAGTGACA	CTCCTAGTCAGGTGGAGCAAAG	657
2	CTGCCTGACATACACCTCCTTC	CACCCTCTCTTCCCACAAATAG	111
3	CCTCCTCTCCGCTTTCCTTCT	CACAAGGCAGCATTGACTGT	192
4	GTTCTTGGTGAGTTCTCCAGTG	GAATGTATGGCAGACCTGGAC	303
5	CTGTCTCAAAGGAGGGAAACAG	CTCCCCTTCTCAATCACACAG	304
6	CACAAGTTCTGCAGCTGTCACT	AGGTAGGGAAGGAAGGAAACAG	346
7	CTACCTCCAGGTCTTGTCCTCTT	CCAGACTTCACTTTGGGACTCT	346
8	GGCCTCCTTTTCATATGCTG	AGAAAGAAAGAGGAGGGAGCAG	261
9	CCTACAAGATCGTCCTCAAACC	CGTCCCATAGGAACAGCAAA	283
10	GAACACCAGCTGCTCATAACTG	GTGCTGGGTGCTGCTAAGATAG	328
11	CAGAAGCTACGGGATGCTAATG	CACCCTCAAAGGAACCACTAAC	207
12	GTCTCACTATGTTGCCCAGCTA	GTGACACAGTCCACCGAGATT	592
13	CCCACACAACACCTAACACAGT	GGTACATCACCGCAATGTGA	286
14	GAACTGCAAACCTCCCTGATAG	GGGGTAGGGAGGACTTCTCTTAT	167
15	CAAGCAGGGATTCTTCTCCTC	GGTCCTGGTCACAGCTTTAAGA	183
16	GGTCCTATTATCAGGTCCTCTCC	GCAGTCAGAGAGAATGGGCTACT	167
17	GGGAAGGGAATGGAGAAAAG	CCACAATAGCCGTTGAGCTT	329^a
18	GCGTATCTTGGTCCTGTGTCTT	ACCAGGAACATCAGCTTCTGAC	553^b

^aPKCβ2 isoform.
^bPKCβ1 isoform.

Table 2. Sequence Variations in the *PRKCB1* Gene in Lung Cancer Cell Lines
Cell Line	Histology	PRKCB1 Coding Region Variants	Exon Number	Amino Acid Change
H23	NSCLC	G702A	7	–
–	–	C1770T	16	–
H125	NSCLC	C79A	1	–
H226	NSCLC	C79A	1	–
–	–	C606T	6	–
H290	NSCLC	T1191C	10	–
H292	NSCLC	C79A	1	–
H322	NSCLC	T1191C	10	–
–	–	C1770T	16	–
H324	NSCLC	C79A	1	–
H358	NSCLC	–	–	–
H520	NSCLC	–	–	–
H522	NSCLC	T1191C	10	–
–	–	C1770T	16	–
H661	NSCLC	–	–	–
EPLC-65H	NSCLC	C79A	1	–
LCLC-97TM1	NSCLC	T1191C	10	–
–	–	C1770T	16	–
LCLC-103H	NSCLC	–	–	–
EPLC-272H	NSCLC	C79A	1	–
A549	NSCLC	–	–	–
H69	SCLC	C79A	1	–
–	–	C119T	1	T40I
H82	SCLC	T1191C	10	–
–	–	C1770T	16	–
H146	SCLC	–	–	–
H209	SCLC	C79A	1	–
H211	SCLC	T1191C	10	–
H417	SCLC	–	–	–
16HC	SCLC	C79A	1	–
–	–	T1191C	10	–
–	–	C1770T	16	–
22H	SCLC	C79A	1	–
–	–	T1191C	10	–
–	–	C1770T	16	–
24H	SCLC	C79A	1	–
–	–	T1191C	10	–
–	–	C1770T	16	–
86M1	SCLC	C79A	1	–
–	–	T1191C	10	–
–	–	C1770T	16	–
DMS76	SCLC	T1191C	10	–
SW210.5	SCLC	C79A	1	–
–	–	C119T	1	T40I
–	–	T1191C	10	–

Abbreviations: NSCLC = non–small-cell lung caner; SCLC = small-cell lung cancer

Table 3. Enzastaurin Concentrations That Induced 50% Inhibition
Cell Line	Proliferation, μM	pPKCβ2 (Ser660),μM	pGSK3β (Ser9), μM	pS6RP (Ser240/244), μM	pAkt (Thr308), μM	pAkt (Ser473), μM	pFKHR (Ser256), μM
H69	0.20	–	–	–	–	–	–
SW210.5	0.07	–	–	–	–	–	–
H211	0.05	–	–	–	–	–	–
DMS79	0.08	–	–	–	–	–	–
H23	0.09	0.08	0.08	0.08	0.08	> 5.00	0.30
H125	0.08	–	–	–	–	–	–
H322	0.08	–	–	–	–	–	–
A549	0.07	–	–	–	0.20	> 5.00	0.20